Sterility:Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in-house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube; see recipe) 1N HCl (500 μl in a microfuge tube) Dulbecco's PBS with Calcium and Magnesium (PBS ++ ) chilled to 4°C and at room temperature MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) 10 and 1000 μl pipettes and tips ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips (these are optional but really worth the money to get consistent results) www.gilson.com/en/Pipette/Products/44.224/Default.aspx#.VIm6raNOmRs Rectangular ice bucket packed with ice Parafilm Chilling/heating dry bath (Echotherm from Torrey Pines Scientific).
Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Materials Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube) 1N HCl (500 μl in a microfuge tube) PBS ++ chilled to 4°C MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips Rectangular ice bucket packed with ice 10 cm tissue culture dish Cell scrapers (3-4: Costar 3008) Aluminum foil 50 mM Tris buffer (see recipe) Scintillation vial or small 50 ml beaker 500 mM acetic acid 20 mM acetic acid chilled to 4°C 2-4 liter beaker Magnetic stir bar to large beaker Magnetic stir bar for scintillation vial 5 mg/ml Atto-488 NHS-ester dye in DMSO (see recipe) 50 mM Borate buffer pH 9.0 (see recipe) 3 ml Slide-A-Lyzer Dialysis cassette (G2) with 20,000 MW cutoff (Pierce: no syringe required) Plastic wrap Sircol Collagen Assay kit (available from accuratechemical.com) Calculating collagen concentrations and volumes list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Calculate reagent volumes for 5 mls of a 3 mg/ml collagen gel (see step 1 of Basic Protocol 1 for sample calculation): list-behavior=simple prefix-word= mark-type=none max-label-size=0 6 mg/ml Rat tail collagen: 2.5 ml 10X DMEM: 250 μl 10X RB: 250 μl 1N NaOH: ∼16 μl PBS ++ : ∼1984 μl Pre-chill all gel components on ice (collagen stock solution, 10X DMEM, 10X RB, 1N NaOH, 1N HCl, PBS ++ ) and the 10 cm tissue culture dish.
Article Title: Single-cell Motility Analysis of Tethered Human Spermatozoa
Article Snippet: Polypropylene sterile sample collection containers (SARSTEDT, catalog number: 75.9922.745) Centrifuge Tubes, 50 ml and 15 ml (Olympus Plastics, catalog numbers: 28-106 and 28-101) Positive Displacement Pipettes and Tips (Gilson, MICROMAN) Micropipettes and Tips (Gilson, Pipetman) 5 mm round coverslips (WPI, catalog number: 502040) 0.22 μm Polyethersulfone (PES) Vacuum-Driven Filter System, 500 ml and 1 L (Olympus Plastics, catalog numbers: 25-227 and 25-229) MLA Pipette Tips, 200 μl (Vista Labs, catalog number: 9025) Osmolarity Calibration Standard (ELITech, OPTIMOLE, catalog number: OA-029) Osmometer Paper Sample Disks (Wescor, VAPRO Sample Disks, catalog number: SS-033) Sodium Chloride (NaCl) (Sigma-Aldrich, catalog number: S7653) Potassium Chloride (KCl) (Fisher Scientific, catalog number: P217) Magnesium Sulfate Heptahydrate (MgSO 4 ·7H 2 O) (Sigma-Aldrich, catalog number: 63140) Monopotassium Phosphate (KH 2 PO 4 ) (Sigma-Aldrich, catalog number: 60216) Calcium Chloride Dihydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: 21097) HEPES (Fisher Scientific, catalog number: BP310) D-(+)-Glucose (C 6 H 12 O 6 ) (Sigma-Aldrich, catalog number: G8270) Sodium Lactate (C 3 H 5 NaO 3 ) (Sigma-Aldrich, catalog number: L7900) Sodium Pyruvate (C 3 H 3 NaO 3 ) (Sigma-Aldrich, catalog number: P2256) Sodium Hydroxide (NaOH) (Sigma-Aldrich, catalog number: 221465) Sodium Bicarbonate (NaHCO 3 ) (Sigma-Aldrich, catalog number: S5761) Fetal Bovine Serum (FBS) (Gibco, catalog number: 16000036) Hydrochloric Acid (HCl) (Fisher Scientific, catalog number: A144S) Human fibrillogenic peptide SEM1(86-107) (Celtek and CPC Scientific, sequence: DLNALHKTTKSQRHLGGSQQLL) pH 4.0 Standard, Red (EMD Chemicals, catalog number: BX1628-1) pH 7.00 Standard, Yellow (Fisher Scientific, catalog number: SB107-500) pH 10.00 Standard, Blue (Fisher Scientific, catalog number: SB115-500) Phosphate-Buffered Saline (PBS) (Fisher Scientific, catalog number: BP665-1) 70% ethanol (Decon Labs, catalog number: V1001) Human Tubal Fluid (HTF) Solution (see Recipes) High-Saline solution (HS) (see Recipes) Capacitation media (see Recipes) Motility disrupting reagents and control reagents (see Recipes)
Transferring:Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in-house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube; see recipe) 1N HCl (500 μl in a microfuge tube) Dulbecco's PBS with Calcium and Magnesium (PBS ++ ) chilled to 4°C and at room temperature MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) 10 and 1000 μl pipettes and tips ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips (these are optional but really worth the money to get consistent results) www.gilson.com/en/Pipette/Products/44.224/Default.aspx#.VIm6raNOmRs Rectangular ice bucket packed with ice Parafilm Chilling/heating dry bath (Echotherm from Torrey Pines Scientific).
Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Materials Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube) 1N HCl (500 μl in a microfuge tube) PBS ++ chilled to 4°C MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips Rectangular ice bucket packed with ice 10 cm tissue culture dish Cell scrapers (3-4: Costar 3008) Aluminum foil 50 mM Tris buffer (see recipe) Scintillation vial or small 50 ml beaker 500 mM acetic acid 20 mM acetic acid chilled to 4°C 2-4 liter beaker Magnetic stir bar to large beaker Magnetic stir bar for scintillation vial 5 mg/ml Atto-488 NHS-ester dye in DMSO (see recipe) 50 mM Borate buffer pH 9.0 (see recipe) 3 ml Slide-A-Lyzer Dialysis cassette (G2) with 20,000 MW cutoff (Pierce: no syringe required) Plastic wrap Sircol Collagen Assay kit (available from accuratechemical.com) Calculating collagen concentrations and volumes list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Calculate reagent volumes for 5 mls of a 3 mg/ml collagen gel (see step 1 of Basic Protocol 1 for sample calculation): list-behavior=simple prefix-word= mark-type=none max-label-size=0 6 mg/ml Rat tail collagen: 2.5 ml 10X DMEM: 250 μl 10X RB: 250 μl 1N NaOH: ∼16 μl PBS ++ : ∼1984 μl Pre-chill all gel components on ice (collagen stock solution, 10X DMEM, 10X RB, 1N NaOH, 1N HCl, PBS ++ ) and the 10 cm tissue culture dish.
Article Title: Single-cell Motility Analysis of Tethered Human Spermatozoa
Article Snippet: Polypropylene sterile sample collection containers (SARSTEDT, catalog number: 75.9922.745) Centrifuge Tubes, 50 ml and 15 ml (Olympus Plastics, catalog numbers: 28-106 and 28-101) Positive Displacement Pipettes and Tips (Gilson, MICROMAN) Micropipettes and Tips (Gilson, Pipetman) 5 mm round coverslips (WPI, catalog number: 502040) 0.22 μm Polyethersulfone (PES) Vacuum-Driven Filter System, 500 ml and 1 L (Olympus Plastics, catalog numbers: 25-227 and 25-229) MLA Pipette Tips, 200 μl (Vista Labs, catalog number: 9025) Osmolarity Calibration Standard (ELITech, OPTIMOLE, catalog number: OA-029) Osmometer Paper Sample Disks (Wescor, VAPRO Sample Disks, catalog number: SS-033) Sodium Chloride (NaCl) (Sigma-Aldrich, catalog number: S7653) Potassium Chloride (KCl) (Fisher Scientific, catalog number: P217) Magnesium Sulfate Heptahydrate (MgSO 4 ·7H 2 O) (Sigma-Aldrich, catalog number: 63140) Monopotassium Phosphate (KH 2 PO 4 ) (Sigma-Aldrich, catalog number: 60216) Calcium Chloride Dihydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: 21097) HEPES (Fisher Scientific, catalog number: BP310) D-(+)-Glucose (C 6 H 12 O 6 ) (Sigma-Aldrich, catalog number: G8270) Sodium Lactate (C 3 H 5 NaO 3 ) (Sigma-Aldrich, catalog number: L7900) Sodium Pyruvate (C 3 H 3 NaO 3 ) (Sigma-Aldrich, catalog number: P2256) Sodium Hydroxide (NaOH) (Sigma-Aldrich, catalog number: 221465) Sodium Bicarbonate (NaHCO 3 ) (Sigma-Aldrich, catalog number: S5761) Fetal Bovine Serum (FBS) (Gibco, catalog number: 16000036) Hydrochloric Acid (HCl) (Fisher Scientific, catalog number: A144S) Human fibrillogenic peptide SEM1(86-107) (Celtek and CPC Scientific, sequence: DLNALHKTTKSQRHLGGSQQLL) pH 4.0 Standard, Red (EMD Chemicals, catalog number: BX1628-1) pH 7.00 Standard, Yellow (Fisher Scientific, catalog number: SB107-500) pH 10.00 Standard, Blue (Fisher Scientific, catalog number: SB115-500) Phosphate-Buffered Saline (PBS) (Fisher Scientific, catalog number: BP665-1) 70% ethanol (Decon Labs, catalog number: V1001) Human Tubal Fluid (HTF) Solution (see Recipes) High-Saline solution (HS) (see Recipes) Capacitation media (see Recipes) Motility disrupting reagents and control reagents (see Recipes)
Sequencing:Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in-house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube; see recipe) 1N HCl (500 μl in a microfuge tube) Dulbecco's PBS with Calcium and Magnesium (PBS ++ ) chilled to 4°C and at room temperature MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) 10 and 1000 μl pipettes and tips ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips (these are optional but really worth the money to get consistent results) www.gilson.com/en/Pipette/Products/44.224/Default.aspx#.VIm6raNOmRs Rectangular ice bucket packed with ice Parafilm Chilling/heating dry bath (Echotherm from Torrey Pines Scientific).
Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Materials Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube) 1N HCl (500 μl in a microfuge tube) PBS ++ chilled to 4°C MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips Rectangular ice bucket packed with ice 10 cm tissue culture dish Cell scrapers (3-4: Costar 3008) Aluminum foil 50 mM Tris buffer (see recipe) Scintillation vial or small 50 ml beaker 500 mM acetic acid 20 mM acetic acid chilled to 4°C 2-4 liter beaker Magnetic stir bar to large beaker Magnetic stir bar for scintillation vial 5 mg/ml Atto-488 NHS-ester dye in DMSO (see recipe) 50 mM Borate buffer pH 9.0 (see recipe) 3 ml Slide-A-Lyzer Dialysis cassette (G2) with 20,000 MW cutoff (Pierce: no syringe required) Plastic wrap Sircol Collagen Assay kit (available from accuratechemical.com) Calculating collagen concentrations and volumes list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Calculate reagent volumes for 5 mls of a 3 mg/ml collagen gel (see step 1 of Basic Protocol 1 for sample calculation): list-behavior=simple prefix-word= mark-type=none max-label-size=0 6 mg/ml Rat tail collagen: 2.5 ml 10X DMEM: 250 μl 10X RB: 250 μl 1N NaOH: ∼16 μl PBS ++ : ∼1984 μl Pre-chill all gel components on ice (collagen stock solution, 10X DMEM, 10X RB, 1N NaOH, 1N HCl, PBS ++ ) and the 10 cm tissue culture dish.
Article Title: Single-cell Motility Analysis of Tethered Human Spermatozoa
Article Snippet: Polypropylene sterile sample collection containers (SARSTEDT, catalog number: 75.9922.745) Centrifuge Tubes, 50 ml and 15 ml (Olympus Plastics, catalog numbers: 28-106 and 28-101) Positive Displacement Pipettes and Tips (Gilson, MICROMAN) Micropipettes and Tips (Gilson, Pipetman) 5 mm round coverslips (WPI, catalog number: 502040) 0.22 μm Polyethersulfone (PES) Vacuum-Driven Filter System, 500 ml and 1 L (Olympus Plastics, catalog numbers: 25-227 and 25-229) MLA Pipette Tips, 200 μl (Vista Labs, catalog number: 9025) Osmolarity Calibration Standard (ELITech, OPTIMOLE, catalog number: OA-029) Osmometer Paper Sample Disks (Wescor, VAPRO Sample Disks, catalog number: SS-033) Sodium Chloride (NaCl) (Sigma-Aldrich, catalog number: S7653) Potassium Chloride (KCl) (Fisher Scientific, catalog number: P217) Magnesium Sulfate Heptahydrate (MgSO 4 ·7H 2 O) (Sigma-Aldrich, catalog number: 63140) Monopotassium Phosphate (KH 2 PO 4 ) (Sigma-Aldrich, catalog number: 60216) Calcium Chloride Dihydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: 21097) HEPES (Fisher Scientific, catalog number: BP310) D-(+)-Glucose (C 6 H 12 O 6 ) (Sigma-Aldrich, catalog number: G8270) Sodium Lactate (C 3 H 5 NaO 3 ) (Sigma-Aldrich, catalog number: L7900) Sodium Pyruvate (C 3 H 3 NaO 3 ) (Sigma-Aldrich, catalog number: P2256) Sodium Hydroxide (NaOH) (Sigma-Aldrich, catalog number: 221465) Sodium Bicarbonate (NaHCO 3 ) (Sigma-Aldrich, catalog number: S5761) Fetal Bovine Serum (FBS) (Gibco, catalog number: 16000036) Hydrochloric Acid (HCl) (Fisher Scientific, catalog number: A144S) Human fibrillogenic peptide SEM1(86-107) (Celtek and CPC Scientific, sequence: DLNALHKTTKSQRHLGGSQQLL) pH 4.0 Standard, Red (EMD Chemicals, catalog number: BX1628-1) pH 7.00 Standard, Yellow (Fisher Scientific, catalog number: SB107-500) pH 10.00 Standard, Blue (Fisher Scientific, catalog number: SB115-500) Phosphate-Buffered Saline (PBS) (Fisher Scientific, catalog number: BP665-1) 70% ethanol (Decon Labs, catalog number: V1001) Human Tubal Fluid (HTF) Solution (see Recipes) High-Saline solution (HS) (see Recipes) Capacitation media (see Recipes) Motility disrupting reagents and control reagents (see Recipes)
Saline:Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in-house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube; see recipe) 1N HCl (500 μl in a microfuge tube) Dulbecco's PBS with Calcium and Magnesium (PBS ++ ) chilled to 4°C and at room temperature MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) 10 and 1000 μl pipettes and tips ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips (these are optional but really worth the money to get consistent results) www.gilson.com/en/Pipette/Products/44.224/Default.aspx#.VIm6raNOmRs Rectangular ice bucket packed with ice Parafilm Chilling/heating dry bath (Echotherm from Torrey Pines Scientific).
Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Materials Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube) 1N HCl (500 μl in a microfuge tube) PBS ++ chilled to 4°C MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips Rectangular ice bucket packed with ice 10 cm tissue culture dish Cell scrapers (3-4: Costar 3008) Aluminum foil 50 mM Tris buffer (see recipe) Scintillation vial or small 50 ml beaker 500 mM acetic acid 20 mM acetic acid chilled to 4°C 2-4 liter beaker Magnetic stir bar to large beaker Magnetic stir bar for scintillation vial 5 mg/ml Atto-488 NHS-ester dye in DMSO (see recipe) 50 mM Borate buffer pH 9.0 (see recipe) 3 ml Slide-A-Lyzer Dialysis cassette (G2) with 20,000 MW cutoff (Pierce: no syringe required) Plastic wrap Sircol Collagen Assay kit (available from accuratechemical.com) Calculating collagen concentrations and volumes list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Calculate reagent volumes for 5 mls of a 3 mg/ml collagen gel (see step 1 of Basic Protocol 1 for sample calculation): list-behavior=simple prefix-word= mark-type=none max-label-size=0 6 mg/ml Rat tail collagen: 2.5 ml 10X DMEM: 250 μl 10X RB: 250 μl 1N NaOH: ∼16 μl PBS ++ : ∼1984 μl Pre-chill all gel components on ice (collagen stock solution, 10X DMEM, 10X RB, 1N NaOH, 1N HCl, PBS ++ ) and the 10 cm tissue culture dish.
Article Title: Single-cell Motility Analysis of Tethered Human Spermatozoa
Article Snippet: Polypropylene sterile sample collection containers (SARSTEDT, catalog number: 75.9922.745) Centrifuge Tubes, 50 ml and 15 ml (Olympus Plastics, catalog numbers: 28-106 and 28-101) Positive Displacement Pipettes and Tips (Gilson, MICROMAN) Micropipettes and Tips (Gilson, Pipetman) 5 mm round coverslips (WPI, catalog number: 502040) 0.22 μm Polyethersulfone (PES) Vacuum-Driven Filter System, 500 ml and 1 L (Olympus Plastics, catalog numbers: 25-227 and 25-229) MLA Pipette Tips, 200 μl (Vista Labs, catalog number: 9025) Osmolarity Calibration Standard (ELITech, OPTIMOLE, catalog number: OA-029) Osmometer Paper Sample Disks (Wescor, VAPRO Sample Disks, catalog number: SS-033) Sodium Chloride (NaCl) (Sigma-Aldrich, catalog number: S7653) Potassium Chloride (KCl) (Fisher Scientific, catalog number: P217) Magnesium Sulfate Heptahydrate (MgSO 4 ·7H 2 O) (Sigma-Aldrich, catalog number: 63140) Monopotassium Phosphate (KH 2 PO 4 ) (Sigma-Aldrich, catalog number: 60216) Calcium Chloride Dihydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: 21097) HEPES (Fisher Scientific, catalog number: BP310) D-(+)-Glucose (C 6 H 12 O 6 ) (Sigma-Aldrich, catalog number: G8270) Sodium Lactate (C 3 H 5 NaO 3 ) (Sigma-Aldrich, catalog number: L7900) Sodium Pyruvate (C 3 H 3 NaO 3 ) (Sigma-Aldrich, catalog number: P2256) Sodium Hydroxide (NaOH) (Sigma-Aldrich, catalog number: 221465) Sodium Bicarbonate (NaHCO 3 ) (Sigma-Aldrich, catalog number: S5761) Fetal Bovine Serum (FBS) (Gibco, catalog number: 16000036) Hydrochloric Acid (HCl) (Fisher Scientific, catalog number: A144S) Human fibrillogenic peptide SEM1(86-107) (Celtek and CPC Scientific, sequence: DLNALHKTTKSQRHLGGSQQLL) pH 4.0 Standard, Red (EMD Chemicals, catalog number: BX1628-1) pH 7.00 Standard, Yellow (Fisher Scientific, catalog number: SB107-500) pH 10.00 Standard, Blue (Fisher Scientific, catalog number: SB115-500) Phosphate-Buffered Saline (PBS) (Fisher Scientific, catalog number: BP665-1) 70% ethanol (Decon Labs, catalog number: V1001) Human Tubal Fluid (HTF) Solution (see Recipes) High-Saline solution (HS) (see Recipes) Capacitation media (see Recipes) Motility disrupting reagents and control reagents (see Recipes)
Control:Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in-house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube; see recipe) 1N HCl (500 μl in a microfuge tube) Dulbecco's PBS with Calcium and Magnesium (PBS ++ ) chilled to 4°C and at room temperature MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) 10 and 1000 μl pipettes and tips ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips (these are optional but really worth the money to get consistent results) www.gilson.com/en/Pipette/Products/44.224/Default.aspx#.VIm6raNOmRs Rectangular ice bucket packed with ice Parafilm Chilling/heating dry bath (Echotherm from Torrey Pines Scientific).
Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Materials Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube) 1N HCl (500 μl in a microfuge tube) PBS ++ chilled to 4°C MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips Rectangular ice bucket packed with ice 10 cm tissue culture dish Cell scrapers (3-4: Costar 3008) Aluminum foil 50 mM Tris buffer (see recipe) Scintillation vial or small 50 ml beaker 500 mM acetic acid 20 mM acetic acid chilled to 4°C 2-4 liter beaker Magnetic stir bar to large beaker Magnetic stir bar for scintillation vial 5 mg/ml Atto-488 NHS-ester dye in DMSO (see recipe) 50 mM Borate buffer pH 9.0 (see recipe) 3 ml Slide-A-Lyzer Dialysis cassette (G2) with 20,000 MW cutoff (Pierce: no syringe required) Plastic wrap Sircol Collagen Assay kit (available from accuratechemical.com) Calculating collagen concentrations and volumes list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Calculate reagent volumes for 5 mls of a 3 mg/ml collagen gel (see step 1 of Basic Protocol 1 for sample calculation): list-behavior=simple prefix-word= mark-type=none max-label-size=0 6 mg/ml Rat tail collagen: 2.5 ml 10X DMEM: 250 μl 10X RB: 250 μl 1N NaOH: ∼16 μl PBS ++ : ∼1984 μl Pre-chill all gel components on ice (collagen stock solution, 10X DMEM, 10X RB, 1N NaOH, 1N HCl, PBS ++ ) and the 10 cm tissue culture dish.
Article Title: Single-cell Motility Analysis of Tethered Human Spermatozoa
Article Snippet: Polypropylene sterile sample collection containers (SARSTEDT, catalog number: 75.9922.745) Centrifuge Tubes, 50 ml and 15 ml (Olympus Plastics, catalog numbers: 28-106 and 28-101) Positive Displacement Pipettes and Tips (Gilson, MICROMAN) Micropipettes and Tips (Gilson, Pipetman) 5 mm round coverslips (WPI, catalog number: 502040) 0.22 μm Polyethersulfone (PES) Vacuum-Driven Filter System, 500 ml and 1 L (Olympus Plastics, catalog numbers: 25-227 and 25-229) MLA Pipette Tips, 200 μl (Vista Labs, catalog number: 9025) Osmolarity Calibration Standard (ELITech, OPTIMOLE, catalog number: OA-029) Osmometer Paper Sample Disks (Wescor, VAPRO Sample Disks, catalog number: SS-033) Sodium Chloride (NaCl) (Sigma-Aldrich, catalog number: S7653) Potassium Chloride (KCl) (Fisher Scientific, catalog number: P217) Magnesium Sulfate Heptahydrate (MgSO 4 ·7H 2 O) (Sigma-Aldrich, catalog number: 63140) Monopotassium Phosphate (KH 2 PO 4 ) (Sigma-Aldrich, catalog number: 60216) Calcium Chloride Dihydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: 21097) HEPES (Fisher Scientific, catalog number: BP310) D-(+)-Glucose (C 6 H 12 O 6 ) (Sigma-Aldrich, catalog number: G8270) Sodium Lactate (C 3 H 5 NaO 3 ) (Sigma-Aldrich, catalog number: L7900) Sodium Pyruvate (C 3 H 3 NaO 3 ) (Sigma-Aldrich, catalog number: P2256) Sodium Hydroxide (NaOH) (Sigma-Aldrich, catalog number: 221465) Sodium Bicarbonate (NaHCO 3 ) (Sigma-Aldrich, catalog number: S5761) Fetal Bovine Serum (FBS) (Gibco, catalog number: 16000036) Hydrochloric Acid (HCl) (Fisher Scientific, catalog number: A144S) Human fibrillogenic peptide SEM1(86-107) (Celtek and CPC Scientific, sequence: DLNALHKTTKSQRHLGGSQQLL) pH 4.0 Standard, Red (EMD Chemicals, catalog number: BX1628-1) pH 7.00 Standard, Yellow (Fisher Scientific, catalog number: SB107-500) pH 10.00 Standard, Blue (Fisher Scientific, catalog number: SB115-500) Phosphate-Buffered Saline (PBS) (Fisher Scientific, catalog number: BP665-1) 70% ethanol (Decon Labs, catalog number: V1001) Human Tubal Fluid (HTF) Solution (see Recipes) High-Saline solution (HS) (see Recipes) Capacitation media (see Recipes) Motility disrupting reagents and control reagents (see Recipes)
Concentration Assay:Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in-house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube; see recipe) 1N HCl (500 μl in a microfuge tube) Dulbecco's PBS with Calcium and Magnesium (PBS ++ ) chilled to 4°C and at room temperature MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) 10 and 1000 μl pipettes and tips ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips (these are optional but really worth the money to get consistent results) www.gilson.com/en/Pipette/Products/44.224/Default.aspx#.VIm6raNOmRs Rectangular ice bucket packed with ice Parafilm Chilling/heating dry bath (Echotherm from Torrey Pines Scientific).
Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Materials Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube) 1N HCl (500 μl in a microfuge tube) PBS ++ chilled to 4°C MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips Rectangular ice bucket packed with ice 10 cm tissue culture dish Cell scrapers (3-4: Costar 3008) Aluminum foil 50 mM Tris buffer (see recipe) Scintillation vial or small 50 ml beaker 500 mM acetic acid 20 mM acetic acid chilled to 4°C 2-4 liter beaker Magnetic stir bar to large beaker Magnetic stir bar for scintillation vial 5 mg/ml Atto-488 NHS-ester dye in DMSO (see recipe) 50 mM Borate buffer pH 9.0 (see recipe) 3 ml Slide-A-Lyzer Dialysis cassette (G2) with 20,000 MW cutoff (Pierce: no syringe required) Plastic wrap Sircol Collagen Assay kit (available from accuratechemical.com) Calculating collagen concentrations and volumes list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Calculate reagent volumes for 5 mls of a 3 mg/ml collagen gel (see step 1 of Basic Protocol 1 for sample calculation): list-behavior=simple prefix-word= mark-type=none max-label-size=0 6 mg/ml Rat tail collagen: 2.5 ml 10X DMEM: 250 μl 10X RB: 250 μl 1N NaOH: ∼16 μl PBS ++ : ∼1984 μl Pre-chill all gel components on ice (collagen stock solution, 10X DMEM, 10X RB, 1N NaOH, 1N HCl, PBS ++ ) and the 10 cm tissue culture dish.
Article Title: Single-cell Motility Analysis of Tethered Human Spermatozoa
Article Snippet: Polypropylene sterile sample collection containers (SARSTEDT, catalog number: 75.9922.745) Centrifuge Tubes, 50 ml and 15 ml (Olympus Plastics, catalog numbers: 28-106 and 28-101) Positive Displacement Pipettes and Tips (Gilson, MICROMAN) Micropipettes and Tips (Gilson, Pipetman) 5 mm round coverslips (WPI, catalog number: 502040) 0.22 μm Polyethersulfone (PES) Vacuum-Driven Filter System, 500 ml and 1 L (Olympus Plastics, catalog numbers: 25-227 and 25-229) MLA Pipette Tips, 200 μl (Vista Labs, catalog number: 9025) Osmolarity Calibration Standard (ELITech, OPTIMOLE, catalog number: OA-029) Osmometer Paper Sample Disks (Wescor, VAPRO Sample Disks, catalog number: SS-033) Sodium Chloride (NaCl) (Sigma-Aldrich, catalog number: S7653) Potassium Chloride (KCl) (Fisher Scientific, catalog number: P217) Magnesium Sulfate Heptahydrate (MgSO 4 ·7H 2 O) (Sigma-Aldrich, catalog number: 63140) Monopotassium Phosphate (KH 2 PO 4 ) (Sigma-Aldrich, catalog number: 60216) Calcium Chloride Dihydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: 21097) HEPES (Fisher Scientific, catalog number: BP310) D-(+)-Glucose (C 6 H 12 O 6 ) (Sigma-Aldrich, catalog number: G8270) Sodium Lactate (C 3 H 5 NaO 3 ) (Sigma-Aldrich, catalog number: L7900) Sodium Pyruvate (C 3 H 3 NaO 3 ) (Sigma-Aldrich, catalog number: P2256) Sodium Hydroxide (NaOH) (Sigma-Aldrich, catalog number: 221465) Sodium Bicarbonate (NaHCO 3 ) (Sigma-Aldrich, catalog number: S5761) Fetal Bovine Serum (FBS) (Gibco, catalog number: 16000036) Hydrochloric Acid (HCl) (Fisher Scientific, catalog number: A144S) Human fibrillogenic peptide SEM1(86-107) (Celtek and CPC Scientific, sequence: DLNALHKTTKSQRHLGGSQQLL) pH 4.0 Standard, Red (EMD Chemicals, catalog number: BX1628-1) pH 7.00 Standard, Yellow (Fisher Scientific, catalog number: SB107-500) pH 10.00 Standard, Blue (Fisher Scientific, catalog number: SB115-500) Phosphate-Buffered Saline (PBS) (Fisher Scientific, catalog number: BP665-1) 70% ethanol (Decon Labs, catalog number: V1001) Human Tubal Fluid (HTF) Solution (see Recipes) High-Saline solution (HS) (see Recipes) Capacitation media (see Recipes) Motility disrupting reagents and control reagents (see Recipes)
Sircol Collagen Assay:Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in-house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube; see recipe) 1N HCl (500 μl in a microfuge tube) Dulbecco's PBS with Calcium and Magnesium (PBS ++ ) chilled to 4°C and at room temperature MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) 10 and 1000 μl pipettes and tips ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips (these are optional but really worth the money to get consistent results) www.gilson.com/en/Pipette/Products/44.224/Default.aspx#.VIm6raNOmRs Rectangular ice bucket packed with ice Parafilm Chilling/heating dry bath (Echotherm from Torrey Pines Scientific).
Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Materials Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube) 1N HCl (500 μl in a microfuge tube) PBS ++ chilled to 4°C MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips Rectangular ice bucket packed with ice 10 cm tissue culture dish Cell scrapers (3-4: Costar 3008) Aluminum foil 50 mM Tris buffer (see recipe) Scintillation vial or small 50 ml beaker 500 mM acetic acid 20 mM acetic acid chilled to 4°C 2-4 liter beaker Magnetic stir bar to large beaker Magnetic stir bar for scintillation vial 5 mg/ml Atto-488 NHS-ester dye in DMSO (see recipe) 50 mM Borate buffer pH 9.0 (see recipe) 3 ml Slide-A-Lyzer Dialysis cassette (G2) with 20,000 MW cutoff (Pierce: no syringe required) Plastic wrap Sircol Collagen Assay kit (available from accuratechemical.com) Calculating collagen concentrations and volumes list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Calculate reagent volumes for 5 mls of a 3 mg/ml collagen gel (see step 1 of Basic Protocol 1 for sample calculation): list-behavior=simple prefix-word= mark-type=none max-label-size=0 6 mg/ml Rat tail collagen: 2.5 ml 10X DMEM: 250 μl 10X RB: 250 μl 1N NaOH: ∼16 μl PBS ++ : ∼1984 μl Pre-chill all gel components on ice (collagen stock solution, 10X DMEM, 10X RB, 1N NaOH, 1N HCl, PBS ++ ) and the 10 cm tissue culture dish.
Article Title: Single-cell Motility Analysis of Tethered Human Spermatozoa
Article Snippet: Polypropylene sterile sample collection containers (SARSTEDT, catalog number: 75.9922.745) Centrifuge Tubes, 50 ml and 15 ml (Olympus Plastics, catalog numbers: 28-106 and 28-101) Positive Displacement Pipettes and Tips (Gilson, MICROMAN) Micropipettes and Tips (Gilson, Pipetman) 5 mm round coverslips (WPI, catalog number: 502040) 0.22 μm Polyethersulfone (PES) Vacuum-Driven Filter System, 500 ml and 1 L (Olympus Plastics, catalog numbers: 25-227 and 25-229) MLA Pipette Tips, 200 μl (Vista Labs, catalog number: 9025) Osmolarity Calibration Standard (ELITech, OPTIMOLE, catalog number: OA-029) Osmometer Paper Sample Disks (Wescor, VAPRO Sample Disks, catalog number: SS-033) Sodium Chloride (NaCl) (Sigma-Aldrich, catalog number: S7653) Potassium Chloride (KCl) (Fisher Scientific, catalog number: P217) Magnesium Sulfate Heptahydrate (MgSO 4 ·7H 2 O) (Sigma-Aldrich, catalog number: 63140) Monopotassium Phosphate (KH 2 PO 4 ) (Sigma-Aldrich, catalog number: 60216) Calcium Chloride Dihydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: 21097) HEPES (Fisher Scientific, catalog number: BP310) D-(+)-Glucose (C 6 H 12 O 6 ) (Sigma-Aldrich, catalog number: G8270) Sodium Lactate (C 3 H 5 NaO 3 ) (Sigma-Aldrich, catalog number: L7900) Sodium Pyruvate (C 3 H 3 NaO 3 ) (Sigma-Aldrich, catalog number: P2256) Sodium Hydroxide (NaOH) (Sigma-Aldrich, catalog number: 221465) Sodium Bicarbonate (NaHCO 3 ) (Sigma-Aldrich, catalog number: S5761) Fetal Bovine Serum (FBS) (Gibco, catalog number: 16000036) Hydrochloric Acid (HCl) (Fisher Scientific, catalog number: A144S) Human fibrillogenic peptide SEM1(86-107) (Celtek and CPC Scientific, sequence: DLNALHKTTKSQRHLGGSQQLL) pH 4.0 Standard, Red (EMD Chemicals, catalog number: BX1628-1) pH 7.00 Standard, Yellow (Fisher Scientific, catalog number: SB107-500) pH 10.00 Standard, Blue (Fisher Scientific, catalog number: SB115-500) Phosphate-Buffered Saline (PBS) (Fisher Scientific, catalog number: BP665-1) 70% ethanol (Decon Labs, catalog number: V1001) Human Tubal Fluid (HTF) Solution (see Recipes) High-Saline solution (HS) (see Recipes) Capacitation media (see Recipes) Motility disrupting reagents and control reagents (see Recipes)
Labeling:Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in-house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube; see recipe) 1N HCl (500 μl in a microfuge tube) Dulbecco's PBS with Calcium and Magnesium (PBS ++ ) chilled to 4°C and at room temperature MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) 10 and 1000 μl pipettes and tips ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips (these are optional but really worth the money to get consistent results) www.gilson.com/en/Pipette/Products/44.224/Default.aspx#.VIm6raNOmRs Rectangular ice bucket packed with ice Parafilm Chilling/heating dry bath (Echotherm from Torrey Pines Scientific).
Article Title: Generation of 3D collagen gels with controlled, diverse architectures
Article Snippet: Materials Rat tail collagen solution dissolved in 20 mM acetic acid (commercial brands are fine, but in house preparations are usually cleaner and polymerize faster) at a concentration greater than 5 mg/ml (6 mg/ml used here) 10X DMEM (see recipe) 10X reconstitution buffer (10X RB; see recipe) 1N NaOH (500 μl in a microfuge tube) 1N HCl (500 μl in a microfuge tube) PBS ++ chilled to 4°C MatTek Dishes (35mm, #1.5 coverslip, 20 mm opening: Part # P35G-1.5-20-C) ColorpHast pH-indicator-strips with pH range 6.5-10 (EMD Biosciences) 10-100 and 1000 μl Gilson positive displacement pipettes and tips Rectangular ice bucket packed with ice 10 cm tissue culture dish Cell scrapers (3-4: Costar 3008) Aluminum foil 50 mM Tris buffer (see recipe) Scintillation vial or small 50 ml beaker 500 mM acetic acid 20 mM acetic acid chilled to 4°C 2-4 liter beaker Magnetic stir bar to large beaker Magnetic stir bar for scintillation vial 5 mg/ml Atto-488 NHS-ester dye in DMSO (see recipe) 50 mM Borate buffer pH 9.0 (see recipe) 3 ml Slide-A-Lyzer Dialysis cassette (G2) with 20,000 MW cutoff (Pierce: no syringe required) Plastic wrap Sircol Collagen Assay kit (available from accuratechemical.com) Calculating collagen concentrations and volumes list-behavior=enumerated prefix-word= mark-type=decimal max-label-size=0 Calculate reagent volumes for 5 mls of a 3 mg/ml collagen gel (see step 1 of Basic Protocol 1 for sample calculation): list-behavior=simple prefix-word= mark-type=none max-label-size=0 6 mg/ml Rat tail collagen: 2.5 ml 10X DMEM: 250 μl 10X RB: 250 μl 1N NaOH: ∼16 μl PBS ++ : ∼1984 μl Pre-chill all gel components on ice (collagen stock solution, 10X DMEM, 10X RB, 1N NaOH, 1N HCl, PBS ++ ) and the 10 cm tissue culture dish.
Article Title: Single-cell Motility Analysis of Tethered Human Spermatozoa
Article Snippet: Polypropylene sterile sample collection containers (SARSTEDT, catalog number: 75.9922.745) Centrifuge Tubes, 50 ml and 15 ml (Olympus Plastics, catalog numbers: 28-106 and 28-101) Positive Displacement Pipettes and Tips (Gilson, MICROMAN) Micropipettes and Tips (Gilson, Pipetman) 5 mm round coverslips (WPI, catalog number: 502040) 0.22 μm Polyethersulfone (PES) Vacuum-Driven Filter System, 500 ml and 1 L (Olympus Plastics, catalog numbers: 25-227 and 25-229) MLA Pipette Tips, 200 μl (Vista Labs, catalog number: 9025) Osmolarity Calibration Standard (ELITech, OPTIMOLE, catalog number: OA-029) Osmometer Paper Sample Disks (Wescor, VAPRO Sample Disks, catalog number: SS-033) Sodium Chloride (NaCl) (Sigma-Aldrich, catalog number: S7653) Potassium Chloride (KCl) (Fisher Scientific, catalog number: P217) Magnesium Sulfate Heptahydrate (MgSO 4 ·7H 2 O) (Sigma-Aldrich, catalog number: 63140) Monopotassium Phosphate (KH 2 PO 4 ) (Sigma-Aldrich, catalog number: 60216) Calcium Chloride Dihydrate (CaCl 2 ·2H 2 O) (Sigma-Aldrich, catalog number: 21097) HEPES (Fisher Scientific, catalog number: BP310) D-(+)-Glucose (C 6 H 12 O 6 ) (Sigma-Aldrich, catalog number: G8270) Sodium Lactate (C 3 H 5 NaO 3 ) (Sigma-Aldrich, catalog number: L7900) Sodium Pyruvate (C 3 H 3 NaO 3 ) (Sigma-Aldrich, catalog number: P2256) Sodium Hydroxide (NaOH) (Sigma-Aldrich, catalog number: 221465) Sodium Bicarbonate (NaHCO 3 ) (Sigma-Aldrich, catalog number: S5761) Fetal Bovine Serum (FBS) (Gibco, catalog number: 16000036) Hydrochloric Acid (HCl) (Fisher Scientific, catalog number: A144S) Human fibrillogenic peptide SEM1(86-107) (Celtek and CPC Scientific, sequence: DLNALHKTTKSQRHLGGSQQLL) pH 4.0 Standard, Red (EMD Chemicals, catalog number: BX1628-1) pH 7.00 Standard, Yellow (Fisher Scientific, catalog number: SB107-500) pH 10.00 Standard, Blue (Fisher Scientific, catalog number: SB115-500) Phosphate-Buffered Saline (PBS) (Fisher Scientific, catalog number: BP665-1) 70% ethanol (Decon Labs, catalog number: V1001) Human Tubal Fluid (HTF) Solution (see Recipes) High-Saline solution (HS) (see Recipes) Capacitation media (see Recipes) Motility disrupting reagents and control reagents (see Recipes)
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