Microscopy & Microtechniques

  • Figure 1: Fluorescence staining of renal epithelial cells infected with Candida albians using Clipmax (PS)

Fluorescence Staining of Renal Epithelial Cells Infected with Candida albicans using Clipmax

Mar 07 2018 Read 532 Times

Clipmax is a combination of a tissue culture flask 10cm2 with the removable standard microscope slide (25mm x 75mm) as a growing surface. It is intended for cell culture, staining, and immunofluorescence as it does not interfere with fluorescent labels directly on the slide. It is also useful for viral and mycoplasma testing, chromosome studies, toxicity tests and immune cytology. The growth area of the slide is TPP opto-mechanically treated resulting in an optimal cell adhesion and proliferation.

After the cell culture the medium chamber with the biocompatible seal ring can easily be detached from the slide. Cells may then be fixed, stained, cover slipped, and filed with the usual convenience, and be used directly for microscopic view or directly for immunocytochemistry and various cell analyses. The transmission of the crystal-clear microscope slide is reported to be possible as of 250nm with a tailing at 280-300nm and stable between 350–400nm until + 750nm (+92% transmission within 400–800nm). The fluoroscopic measurements are better than with glass, PC, or PS and the protein absorption is less than using glass slides. The thermal and optical features of the Clipmax slide are better than those made from PC, PET or TAC and in addition the slide is resistant to numerous chemicals such as Xylolite, alcohol, ether, (HCl, HNO3, H2SO4), (NaOH, NH4OH) or even H2O2. Also, biological coatings, i.g. with collagen, result in no loss of measuring signals. Finally, the handling of the Clipmax slide is safe because there is no risk of injury, as it is plastic and not glass.

Tony Pawlik from the Hans Knöll Institute Jena cultured a renal epithelial cell line A498 (DSMZ no. ACC 55) to study the interaction between Candida albicans (human pathogenic fungi) and this cell line using the Clipmax tissue culture device. To determine the adhesion rate of the fungi and the invasion rate into the renal epithelial cells it was done by fluorescence staining. The renal epithelial cells grow for 1-2 days to a confluent monolayer. After a medium change the cells are infected by Candia albicans which needs 1-4 hours depending on the assay. After several washing steps with PBS the cells are fixed with Histofix and stained with fluorescence dyes (see Figure 1). The cell line A498 is known to grow very well on all opto-mechanically treated PS surfaces, but not on glass surfaces even when treated with collagen-I, poly-D-lysine or concanavalin A. The cells started to adhere but after the first washing step the cells are washed of.

Conclusion: The A498 cell line adheres very well on the surface of the Clipmax slide and this did not change after several washing steps. The picture below shows the renal epithelial cells using transmitted light microscopy and the Candida cells stained with fluorescence brightener.

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