Cryostat thin sections

IGSS may also be performed on cryostat sections, which may then be plastic embedded for semi-thin sectioning (35)... 

Thin sections for transmission electron microscopy were dry cut on a Reichert Ultracut ultramicrotome with a FC4D cryostatic unit temperatures were -100°C for the specimen and -80°C for the diamond knife. Sections stained with RuO vapor as well as unstained sections were viewed by means of a Zeiss EMIO electron microscope. 

For immunocytochemistry, fixed tissue must be cut in thin sections to be viewed in the light or fluorescence microscope. There are two common ways of sectioning tissue - the cryostat for fixed frozen tissue and the rotary microtome for paraffin-embedded tissue. In animal research, select the sectioning method based on the experimental design. The method that gives the most reliable results and is the simplest should be selected. For immunocytochemistry, the cryostat is a very efficient and reliable method. The rotary microtome of paraffin-embedded material is more complex and problematic. For immunocytochemistry in animal research, the cryostat method is recommended for reasons discussed in this chapter. 

Mount the block on the cryostat head and cut thin sections (10-30 J,m), picking them up onto PLL-coated slides see Notes 1, 5, and 7). 

FIGURE 3.28 Cutting thin sections with cryostat. 1. Cutting tissues into optimal tissue thickness. 2. Keep the slice under the antiroll for a while, until it no longer curls. 3 and 4.Put the indium tin oxide (ITO)-coated glass slides into the chamber, immediately place the slides over the tissue slice, and paste tissues on it. 

In typical DESI imaging experiments, the collected tissue sample is flash frozen in liquid nitrogen and subsequently cut in micron thin sections using a cryostat-microtome the thin tissue slices are thaw mounted onto glass microscope slides for analysis the ions generated by the DESI (Chapter 2) are transported fi om surfaces to the gas phase for mass analysis as the surface is being moved in order to cover the entire sample area a mass spectrum is acquired for each pixel on the surface and finally, tissue images are constructed to display the spatial intensity distribution of individual selected lipid ion. 

Tissue sectioning Set cryostat at -16 C and slice brain or desired tissue into sections 16—20 pm thin. Mount sections on poly-L-lysine coated or positively charged slides. Allow sections to dry overnight at room temperature. Tissue at this stage (fresh frozen, unfixed) should be stored at -20 C. 

Cryostat sections should be prepared as thin as possible, 4-8 pm in thickness. Air-dry slides for 1-3 h at room temperature (see Note 7). 

Cryostat sections should be prepared as thin as possible, 4-8 pm in thickness. 

Carrots treated with the dye-biopolymer complexes and processed under the optimum condition as determined by RSM were rehydrated and fixed for light microscopic examination. Thin slices of carrots (2-3 mm) were fixed on cork board and frozen in isopentane at -160°C for 15 sec. The frozen samples were warmed up to -20°C in a histostat cryostat microtome chamber and cut into sections of 28 micron thickness. The sections were fixed on slides with glycerol gel and dried in an oven at 35°C to remove any air bubble under the slides. The slides were examined and photographed under a light microscope at 500 and 785 times magnification. 

For EL-. (7-, and C7ph-detected resonances (see Section 1.4.3), thin copper wire leads were inserted into the quartz dewar of the lie gas flow cryostat to provide bias. The cr- and cTpu-detected resonances were recorded by measuring the microwave-induced changes in the current resulting in similar changes in the voltage across a standard resistor connected in series with the LED. 

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