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intellesis trainable segmentation module  (Carl Zeiss)


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    Carl Zeiss intellesis trainable segmentation module
    A WT or TAp73 KO H1299 cells were transfected with the indicated plasmids for 24 h and IF carried out against ATP5B (green) with DAPI nuclear counterstain (blue). Cells transfected with HA-TAp73α expression plasmid were stained for HA as a transfection control (red). Lower magnification images, scale bar = 10 μm; Callout images, scale bar = 4 μm. B Representative western blot of OPA1 expression following transfection of WT and TAp73 KO cells with pCMW-OPA1 construct. C Quantification of mitochondrial morphology from ( A ) using Zeiss <t>Intellesis</t> module, trained to segment individual mitochondria. Statistical significance for each condition was calculated using Student’s t-test, comparing with WT EV control (column 1); *p < 0.05, (ns) not significant (n = 3). D Transmission electron micrographs of mitochondrial morphology from WT and TAp73 KO cells. Scale bar = 100 nm. E Mitochondrial length measurements obtained from ( D ). ****p < 0.0001 in Student’s t-test. A minimum of 100 mitochondria were measured from n = 3 independent biological replicates. F , G Mitochondrial stress test performed on Seahorse XFe96 analyser. Canonical mitochondrial inhibitors injected sequentially as labelled (Oligomycin = 2 μM, FCCP = 500 nM, Antimycin A/ Rotenone = 2 μM). The indicated mitochondrial stress test parameters were calculated from OCR data. Data were corrected for non-mitochondrial OCR, normalised to cell number, and are shown as mean ± SD (n = 3). *P ≤ 0.05 and **P ≤ 0.01 in Student’s t-test relative to WT control. H Western blot of the indicated ETC subunits in wild-type and TAp73 KO cells, obtained using OXPHOS antibody cocktail. I qPCR against mt-CO2 , expressed relative to expression of nuclear encoded β2-microglobulin . Relative expression was calculated using the ΔΔCt method and expressed as a percentage of wild-type control (n = 2 biological replicates, each tested in triplicate). n.s not significant in Student’s t-test.
    Intellesis Trainable Segmentation Module, supplied by Carl Zeiss, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/product/intellesis+trainable+segmentation+module/pmc11549358-110-10-14?v=Carl+Zeiss
    Average 90 stars, based on 1 article reviews
    intellesis trainable segmentation module - by Bioz Stars, 2026-08
    90/100 stars

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    1) Product Images from "TAp73 regulates mitochondrial dynamics and multiciliated cell homeostasis through an OPA1 axis"

    Article Title: TAp73 regulates mitochondrial dynamics and multiciliated cell homeostasis through an OPA1 axis

    Journal: Cell Death & Disease

    doi: 10.1038/s41419-024-07130-6

    A WT or TAp73 KO H1299 cells were transfected with the indicated plasmids for 24 h and IF carried out against ATP5B (green) with DAPI nuclear counterstain (blue). Cells transfected with HA-TAp73α expression plasmid were stained for HA as a transfection control (red). Lower magnification images, scale bar = 10 μm; Callout images, scale bar = 4 μm. B Representative western blot of OPA1 expression following transfection of WT and TAp73 KO cells with pCMW-OPA1 construct. C Quantification of mitochondrial morphology from ( A ) using Zeiss Intellesis module, trained to segment individual mitochondria. Statistical significance for each condition was calculated using Student’s t-test, comparing with WT EV control (column 1); *p < 0.05, (ns) not significant (n = 3). D Transmission electron micrographs of mitochondrial morphology from WT and TAp73 KO cells. Scale bar = 100 nm. E Mitochondrial length measurements obtained from ( D ). ****p < 0.0001 in Student’s t-test. A minimum of 100 mitochondria were measured from n = 3 independent biological replicates. F , G Mitochondrial stress test performed on Seahorse XFe96 analyser. Canonical mitochondrial inhibitors injected sequentially as labelled (Oligomycin = 2 μM, FCCP = 500 nM, Antimycin A/ Rotenone = 2 μM). The indicated mitochondrial stress test parameters were calculated from OCR data. Data were corrected for non-mitochondrial OCR, normalised to cell number, and are shown as mean ± SD (n = 3). *P ≤ 0.05 and **P ≤ 0.01 in Student’s t-test relative to WT control. H Western blot of the indicated ETC subunits in wild-type and TAp73 KO cells, obtained using OXPHOS antibody cocktail. I qPCR against mt-CO2 , expressed relative to expression of nuclear encoded β2-microglobulin . Relative expression was calculated using the ΔΔCt method and expressed as a percentage of wild-type control (n = 2 biological replicates, each tested in triplicate). n.s not significant in Student’s t-test.
    Figure Legend Snippet: A WT or TAp73 KO H1299 cells were transfected with the indicated plasmids for 24 h and IF carried out against ATP5B (green) with DAPI nuclear counterstain (blue). Cells transfected with HA-TAp73α expression plasmid were stained for HA as a transfection control (red). Lower magnification images, scale bar = 10 μm; Callout images, scale bar = 4 μm. B Representative western blot of OPA1 expression following transfection of WT and TAp73 KO cells with pCMW-OPA1 construct. C Quantification of mitochondrial morphology from ( A ) using Zeiss Intellesis module, trained to segment individual mitochondria. Statistical significance for each condition was calculated using Student’s t-test, comparing with WT EV control (column 1); *p < 0.05, (ns) not significant (n = 3). D Transmission electron micrographs of mitochondrial morphology from WT and TAp73 KO cells. Scale bar = 100 nm. E Mitochondrial length measurements obtained from ( D ). ****p < 0.0001 in Student’s t-test. A minimum of 100 mitochondria were measured from n = 3 independent biological replicates. F , G Mitochondrial stress test performed on Seahorse XFe96 analyser. Canonical mitochondrial inhibitors injected sequentially as labelled (Oligomycin = 2 μM, FCCP = 500 nM, Antimycin A/ Rotenone = 2 μM). The indicated mitochondrial stress test parameters were calculated from OCR data. Data were corrected for non-mitochondrial OCR, normalised to cell number, and are shown as mean ± SD (n = 3). *P ≤ 0.05 and **P ≤ 0.01 in Student’s t-test relative to WT control. H Western blot of the indicated ETC subunits in wild-type and TAp73 KO cells, obtained using OXPHOS antibody cocktail. I qPCR against mt-CO2 , expressed relative to expression of nuclear encoded β2-microglobulin . Relative expression was calculated using the ΔΔCt method and expressed as a percentage of wild-type control (n = 2 biological replicates, each tested in triplicate). n.s not significant in Student’s t-test.

    Techniques Used: Transfection, Expressing, Plasmid Preparation, Staining, Control, Western Blot, Construct, Transmission Assay, Injection



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    A WT or TAp73 KO H1299 cells were transfected with the indicated plasmids for 24 h and IF carried out against ATP5B (green) with DAPI nuclear counterstain (blue). Cells transfected with HA-TAp73α expression plasmid were stained for HA as a transfection control (red). Lower magnification images, scale bar = 10 μm; Callout images, scale bar = 4 μm. B Representative western blot of OPA1 expression following transfection of WT and TAp73 KO cells with pCMW-OPA1 construct. C Quantification of mitochondrial morphology from ( A ) using Zeiss <t>Intellesis</t> module, trained to segment individual mitochondria. Statistical significance for each condition was calculated using Student’s t-test, comparing with WT EV control (column 1); *p < 0.05, (ns) not significant (n = 3). D Transmission electron micrographs of mitochondrial morphology from WT and TAp73 KO cells. Scale bar = 100 nm. E Mitochondrial length measurements obtained from ( D ). ****p < 0.0001 in Student’s t-test. A minimum of 100 mitochondria were measured from n = 3 independent biological replicates. F , G Mitochondrial stress test performed on Seahorse XFe96 analyser. Canonical mitochondrial inhibitors injected sequentially as labelled (Oligomycin = 2 μM, FCCP = 500 nM, Antimycin A/ Rotenone = 2 μM). The indicated mitochondrial stress test parameters were calculated from OCR data. Data were corrected for non-mitochondrial OCR, normalised to cell number, and are shown as mean ± SD (n = 3). *P ≤ 0.05 and **P ≤ 0.01 in Student’s t-test relative to WT control. H Western blot of the indicated ETC subunits in wild-type and TAp73 KO cells, obtained using OXPHOS antibody cocktail. I qPCR against mt-CO2 , expressed relative to expression of nuclear encoded β2-microglobulin . Relative expression was calculated using the ΔΔCt method and expressed as a percentage of wild-type control (n = 2 biological replicates, each tested in triplicate). n.s not significant in Student’s t-test.
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    A WT or TAp73 KO H1299 cells were transfected with the indicated plasmids for 24 h and IF carried out against ATP5B (green) with DAPI nuclear counterstain (blue). Cells transfected with HA-TAp73α expression plasmid were stained for HA as a transfection control (red). Lower magnification images, scale bar = 10 μm; Callout images, scale bar = 4 μm. B Representative western blot of OPA1 expression following transfection of WT and TAp73 KO cells with pCMW-OPA1 construct. C Quantification of mitochondrial morphology from ( A ) using Zeiss Intellesis module, trained to segment individual mitochondria. Statistical significance for each condition was calculated using Student’s t-test, comparing with WT EV control (column 1); *p < 0.05, (ns) not significant (n = 3). D Transmission electron micrographs of mitochondrial morphology from WT and TAp73 KO cells. Scale bar = 100 nm. E Mitochondrial length measurements obtained from ( D ). ****p < 0.0001 in Student’s t-test. A minimum of 100 mitochondria were measured from n = 3 independent biological replicates. F , G Mitochondrial stress test performed on Seahorse XFe96 analyser. Canonical mitochondrial inhibitors injected sequentially as labelled (Oligomycin = 2 μM, FCCP = 500 nM, Antimycin A/ Rotenone = 2 μM). The indicated mitochondrial stress test parameters were calculated from OCR data. Data were corrected for non-mitochondrial OCR, normalised to cell number, and are shown as mean ± SD (n = 3). *P ≤ 0.05 and **P ≤ 0.01 in Student’s t-test relative to WT control. H Western blot of the indicated ETC subunits in wild-type and TAp73 KO cells, obtained using OXPHOS antibody cocktail. I qPCR against mt-CO2 , expressed relative to expression of nuclear encoded β2-microglobulin . Relative expression was calculated using the ΔΔCt method and expressed as a percentage of wild-type control (n = 2 biological replicates, each tested in triplicate). n.s not significant in Student’s t-test.

    Journal: Cell Death & Disease

    Article Title: TAp73 regulates mitochondrial dynamics and multiciliated cell homeostasis through an OPA1 axis

    doi: 10.1038/s41419-024-07130-6

    Figure Lengend Snippet: A WT or TAp73 KO H1299 cells were transfected with the indicated plasmids for 24 h and IF carried out against ATP5B (green) with DAPI nuclear counterstain (blue). Cells transfected with HA-TAp73α expression plasmid were stained for HA as a transfection control (red). Lower magnification images, scale bar = 10 μm; Callout images, scale bar = 4 μm. B Representative western blot of OPA1 expression following transfection of WT and TAp73 KO cells with pCMW-OPA1 construct. C Quantification of mitochondrial morphology from ( A ) using Zeiss Intellesis module, trained to segment individual mitochondria. Statistical significance for each condition was calculated using Student’s t-test, comparing with WT EV control (column 1); *p < 0.05, (ns) not significant (n = 3). D Transmission electron micrographs of mitochondrial morphology from WT and TAp73 KO cells. Scale bar = 100 nm. E Mitochondrial length measurements obtained from ( D ). ****p < 0.0001 in Student’s t-test. A minimum of 100 mitochondria were measured from n = 3 independent biological replicates. F , G Mitochondrial stress test performed on Seahorse XFe96 analyser. Canonical mitochondrial inhibitors injected sequentially as labelled (Oligomycin = 2 μM, FCCP = 500 nM, Antimycin A/ Rotenone = 2 μM). The indicated mitochondrial stress test parameters were calculated from OCR data. Data were corrected for non-mitochondrial OCR, normalised to cell number, and are shown as mean ± SD (n = 3). *P ≤ 0.05 and **P ≤ 0.01 in Student’s t-test relative to WT control. H Western blot of the indicated ETC subunits in wild-type and TAp73 KO cells, obtained using OXPHOS antibody cocktail. I qPCR against mt-CO2 , expressed relative to expression of nuclear encoded β2-microglobulin . Relative expression was calculated using the ΔΔCt method and expressed as a percentage of wild-type control (n = 2 biological replicates, each tested in triplicate). n.s not significant in Student’s t-test.

    Article Snippet: Mitochondrial fragmentation was quantified in an unbiased assay, using the Intellesis trainable segmentation module (Zeiss), which quantified mean mitochondrial area across biological replicates (Fig. ).

    Techniques: Transfection, Expressing, Plasmid Preparation, Staining, Control, Western Blot, Construct, Transmission Assay, Injection