Poly d lysine coated fluorodish

Manufactured by World Precision Instruments

The Poly-D-lysine coated FluoroDish is a specialized cell culture dish designed for fluorescence microscopy applications. The dish features a glass bottom coated with poly-D-lysine, a positively charged polymer that promotes cell attachment and growth. The dish is suitable for a variety of cell types and can be used to study cellular processes under fluorescence imaging.

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Lab products found in correlation

Zen 2, by Zeiss (4 mentions) Spinning disc confocal microscope, by Zeiss (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions) Collagenase, by Merck Group (1 mentions) Tnb 100 lipid protein complex, by Harvard Bioscience (1 mentions) Nerve growth factor 7s, by Alomone (1 mentions)

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FluoroDish (125 citations) FluoroDishes (35 citations)

5 protocols using poly d lysine coated fluorodish

Asymmetric GSC Division Dynamics

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To examine the temporal dynamics of cellular processes during asymmetric GSC divisions, we conducted live cell imaging with high temporal resolution (e.g. 30sec, 60sec or 5min interval as mention in the figure legend and supplemental movie legend). To perform live cell imaging, adult Drosophila testes were dissected in a medium containing Schneider’s insect medium with 200 μg/ml insulin, 15% (vol/vol) FBS, 0.6x pen/strep, with pH value at approximately 7.0, which we called “live cell medium”. Testes were then placed on a Poly-D-lysine coated FluoroDish (World Precision Instrument, Inc.), which contains the live cell medium as described.
All movies were taken using spinning disc confocal microscope (Zeiss) equipped with an evolve^−™ camera (Photometrics), using a 63x Zeiss objective (1.4 NA) at 29°C. The ZEN 2 software (Zeiss) was used for acquisition with 2×2 binning. Mitotic cells were used to reconstruct 3-D movies using Imaris software (Bitplane). All videos for live cells are shown in Movie S1 – Movie S7 and Movie S13.

Ranjan R., Snedeker J, & Chen X. (2019). Asymmetric centromeres differentially coordinate with mitotic machinery to ensure biased sister chromatid segregation in germline stem cells. Cell stem cell, 25(5), 666-681.e5.

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Live Cell Imaging of Asymmetric GSC Divisions

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All live cell imaging experiments were performed before cell cycle arrest, as detailed in Ranjan and Chen (2021) (link). To examine the temporal dynamics of cellular processes during asymmetric GSC divisions, we conducted live cell imaging with high temporal resolution (e.g. 1min or 2min interval as mentioned in the supplemental movie legends). To perform live cell imaging, adult Drosophila testes were dissected in a medium containing Schneider’s medium with 200μg/ml insulin, 15% (vol/vol) FBS, 0.6x pen/strep, with pH value at approximately 7.0, which we call “live cell medium” as reported previously (Ranjan et al., 2019 (link)). Testes were then placed on a Poly-D-lysine coated FluoroDish (World Precision Instrument, Inc.), which contains the live cell medium as described. All movies were taken using spinning disc confocal microscope (Zeiss) equipped with an evolve^-™ camera (Photometrics), using a 63× objective (1.4 NA) at 29°C. The ZEN 2 software (Zeiss) is used for acquisition with 2x2 binning. Representative videos for live cells are shown in Videos S1, S2, S3, S4, S5, S6, and S7. Related to Figures 1, 2, 5, and S5.

Ranjan R., Snedeker J., Wooten M., Chu C., Bracero S., Mouton T, & Chen X. (2022). Differential condensation of sister chromatids acts with Cdc6 to ensure asynchronous S-phase entry in Drosophila male germline stem cell lineage. Developmental cell, 57(9), 1102-1118.e7.

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Live Cell Imaging of Histone Dynamics in Drosophila Germline Stem Cells

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Live cell imaging was performed between 12 and 30 hours after heat shock treatment, as detailed in [94 (link)]. To examine the inheritance pattern of histones during asymmetric GSC divisions, we conducted live cell imaging with high temporal resolution (e.g., 5-minute interval as mention in the S1–S5 Movie legends). To perform live cell imaging, adult Drosophila testes were dissected in a “live cell medium” as reported previously [65 (link)]. Live cell medium contains Schneider’s insect medium with 200 μg/ml insulin, 15% (vol/vol) fetal bovine serum (FBS), 0.6× pen/strep, with pH value at approximately 7.0. Testes were then placed on a Poly-D-lysine coated FluoroDish (World Precision Instrument), which contains the live cell medium as described. All movies were taken using spinning disc confocal microscope (Zeiss) equipped with an evolve camera (Photometrics), using a 63× Zeiss objective (1.4 NA) at 29°C. The ZEN 2 software (Zeiss) was used for acquisition with 2 × 2 binning. All videos for live cells are shown in S1–S5 Movies. WT histone H3 and mutant H3A31S movies were acquired between 22 and 30 hours post heat shock treatment. WT H3.3 and mutant H3.3S31A movies were acquired between 12 and 25 hours post heat shock treatment.

Chandrasekhara C., Ranjan R., Urban J.A., Davis B.E., Ku W.L., Snedeker J., Zhao K, & Chen X. (2023). A single N-terminal amino acid determines the distinct roles of histones H3 and H3.3 in the Drosophila male germline stem cell lineage. PLOS Biology, 21(5), e3002098.

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Trigeminal Ganglion Cell Isolation

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Cultures were performed as previously described (7 (link)). Five days after surgery, mice were euthanized, and the TGs were removed. For each experiment, ganglia of two or three animals were combined, transferred in sterile DMEM (Thermo Fisher Scientific), and fragmented. For cell dissociation, TG fragments were incubated in a mix of protease (Sigma-Aldrich; 1:2000) and collagenase (Sigma-Aldrich; 1:1000) in DMEM at 37°C and 5% CO₂ for 45 min. The cell suspension was then washed in DMEM and TNB 100 medium (Biochrom GmbH), supplemented with TNB 100 lipid-protein complex (Biochrom GmbH), streptomycin/penicillin (100 μg/ml; Thermo Fisher Scientific), and 1 nM nerve growth factor-7S (Alomone Labs), and cell clusters were triturated. Twenty microliters of cell suspension was applied per poly-d-lysine–coated FluoroDish (World Precision Instruments) and incubated in 1-ml TNB medium for 15 to 18 hours at 37°C and 5% CO₂.

Bernal L., Sotelo-Hitschfeld P., König C., Sinica V., Wyatt A., Winter Z., Hein A., Touska F., Reinhardt S., Tragl A., Kusuda R., Wartenberg P., Sclaroff A., Pfeifer J.D., Ectors F., Dahl A., Freichel M., Vlachova V., Brauchi S., Roza C., Boehm U., Clapham D.E., Lennerz J.K, & Zimmermann K. (2021). Odontoblast TRPC5 channels signal cold pain in teeth. Science Advances, 7(13), eabf5567.

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Live Cell Imaging of Asymmetric GSC Divisions

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