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The physical properties of the EVA film allow absorption of energy from the IR laser while permitting wavelengths of the visible spectrum to pass through for microscopic visualization of cryosections. The cells or tissue below the applied IR laser reaches peak temperatures of 90��C for http://www.selleck.cn/products/wortmannin.html to the LCM procedure. The ��sandwich�� structure protects the tissue from contamination, and the compatible CapSure Macro cap http://www.selleckchem.com/products/LY294002.html without rails allows for a larger capture area and cap placement directly on the flat PEN membrane, thereby eliminating laser focusing problems caused by tilting of cap in thin corneal specimens. Finally, the elimination of both traditional sheer forces and substratum forces by UV cutting permits a capture efficiency of 100%. The improved efficiency reduces the capture time and supports maintenance of biomolecule integrity. Currently, the more precise immuno-LCM (Espina et?al. 2007) is not feasible in limbal research combining LCM with RNA sequencing due to the adverse effect of staining protocols on RNA integrity, increased time for cell capture and lack of specific biomarkers for LESCs. The advantages of LCM are many. The technique is fast, thereby allowing procurement of cells with intact biomolecules. Expression profiles are retained in the captured cell populations, if tissue blocks are flash-frozen directly upon removal from donors (Spruessel et?al. 2004). The use of sterile caps and PEN membrane slides allows capture of cells http://www.selleckchem.com/products/CHIR-99021.html without contamination, which is especially important as downstream analyses become increasingly more sensitive (NGS; Emmert-Buck et?al. 1996). Finally, it is possible to visually inspect both the cap and the tissue section after capture for quality control. As previously stated (Bonner et?al. 1997), the LCM procedure only affects cells to be captured, rendering surrounding cells fully accessible for further capture, which is very important when performing comparative molecular analysis of adjacent cell populations. Laser capture microdissection is, though, very demanding from a technical point of view, when downstream applications require intact biomolecules such as RNA. First, the tissue must be flash-frozen to retain biomolecules in their original state.