Tms f light microscope

After the last 4^th passage single cells were plated at a density of 3 × 10⁴ cells/ml or after irradiation at a density of 5 × 10⁴ cells/ml in an uncoated μ-Slide 8 well (iBidi, GmbH, Martinsried, Germany). Representative pictures for each n were taken in 4 consistent regions of each well using a Leica DFC 295 camera and a Nikon TMS-F light microscope. For the irradiation experiment a Nikon ECLIPSE TE20 microscope with an INFINITY1–2C camera and Infinite analyze software were used to capture the pictures.
The diameter of each sphere was measured and the corresponding volume (in μm³) of the sphere was calculated offline for the 24 and 48 hour time using Image J 1.45s (Wayne Rasband, NIH, USA).

The fixed tissues on the slides were immersed in Xylene to remove the wax. The dewaxed samples were passed through a series of ethanol (absolute ethanol, 90 and 70 %). After this treatment, the samples were stained in safranin for 10 min, washed again in a series of ethanol (50, 70 % and absolute) and counterstained in Fast green for 1 min. The samples were then cleared in clove oil and mounted in Euparol (Flatters and Garvett Ltd., England). Examination of starch cells were done using a TMS-F light Microscope and photomicrographs of the slide taken using a Nikon camera (Nikon Co., Tokyo, Japan) attached to the microscope at a magnification of 145.

Dried, ground samples were used for the examination of the starch granules. A minute quality of the sample was added to a small drop of water on a slide and thoroughly mixed taking care not to break any air bubbles. The mixture is then covered with cover slip. Excess water was removed by means of a filter paper and a little dilute iodine was run under the cover slip. Microscopical examination was done using a TMS-F Light Microscope and photomicrographs of the slide taken using a Nikon camera (Nikon Co., Tokyo, Japan) attached to the microscope at a magnification of 145.