Journal: Nature methods

Article Title: Live-cell super-resolved PAINT imaging of pN cellular traction forces

doi: 10.1038/s41592-020-0929-2

Figure Lengend Snippet: (a) Live-cell sf-tPAINT imaging of platelet tension displayed using different time windows (ranging from 50 sec to 1000 sec). The apparent length or width of cellular tension features depends on the number of frames that are integrated to produce a super-resolved image. To demonstrate this point, we rendered the lamellipodial edge of 3 human platelets (from n = 3 independent experiments, 2 platelets shown) and measured the apparent width of the lamellipodial edge tension ring as tPAINT data is integrated over various time windows. Super-resolved tPAINT images were rendered as greyscale images, and ring width was measured via linescan analysis (black dots). The data were fit to a gaussian via Matlab’s curvefitting tool (blue line). The measured FWHM of the fitted gaussians depends on the number of frames integrated to produce the super-resolved tPAINT image. (b) Plot showing that the localization density generally increased with increasing the number of integrated frames. Each color shows a unique ROI. (c) Plot showing the relation between the FWHM of the tension ring and the number of integrated frames. The data shown are from 3 human platelets from n = 3 independent experiments (2 linescans per platelet). In principle, it is desirable to use the minimum number of frames possible to render an image in order to minimize feature blurring due to cellular dynamics during the imaging window; however, image quality decreases, with localizations becoming more punctate, when fewer frames are integrated. To produce high-quality tPAINT images, these considerations must both be balanced. All scale bars are 2 μm.

Article Snippet: The data were fit to a gaussian via Matlab’s curvefitting tool (blue line).

Techniques: Imaging

Journal: Journal of visualized experiments : JoVE

Article Title: Direct-Coupled Electroretinogram (DC-ERG) for Recording the Light-Evoked Electrical Responses of the Mouse Retinal Pigment Epithelium

doi: 10.3791/61491

Figure Lengend Snippet: Table of Materials

Article Snippet: Maintain at RT In-Line Filter Whatman 6722–5001 To protect vacuum pump from aerosols Low Noise Cable for Microelectrode Holders WPI Inc 5372 Suggested for improving the length and placement of the cables and electrode holder assemblies Magnetic Ball Joint WPI Inc 500871 For magnetically positioning the electrode holder assembly on the stage MatLab Mathworks MatLab: For editing the analysis software MatLab Curvefit Toolbox Mathworks Toolbox for MatLab (only required for editing the analysis software) MatLab Compiler Mathworks Toolbox for MatLab (only required for editing and re-releasing the analysis software) MatLab Runtime version 9.5 Mathworks R2018b (9.5) Required to run the analysis software: https://www.mathworks.com/products/compiler/matlab-runtime.html Microelectrode Holders (45 degrees) WPI Inc MEH345–15 For holding the capillaries Needle (25 ga) Covidien 8881250313 For filling the capillary tubes with HBSS Needle (ground) Electrode Rhythmlink 13mm - one elctrode Subdermal needle electrode (ground) for mouse (13mm long, 0.4mm diameter needle, 1.5m leadwire) Regulator/Power Conditioner Furman P-1800 Or equivalent to remove DC-offset from noise introduced through power line Syringe (12 mL) Monoject 1181200777 For filling the capillary tubes with HBSS T-clip Cole-Parmer 06852–20 For electrode holder assembly Vacuum Desiccator Bel-Art 420120000 Clear polycarbonate bottom & cover Pharmacological treatment Lubricant eye gel Alcon 0078–0429-47 Helps lubricate corneal surface and maintain electrical contact with capillary electrodes Phenylephrine Hydrochloride 2.5% Akorn 17478–201-15 Short acting mydriatic eye drops (for pupil dilation) Proparacaine Hydrochloride 0.5% Akorn 17478–263-12 Local anesthetic for ophthalmic instillation Tropicamide 0.5% Akorn 17478–101-12 Short acting mydriatic eye drops (for pupil dilation) Xylazine AnaSed sc-362949Rx Analgesic and muscle relaxant Zetamine (Ketamine HCl) VetOne 501072 Anesthetic for intramuscular injections Open in a separate window list-behavior=simple prefix-word= mark-type=none max-label-size=4 1.1.

Techniques: Convection, Software