Leica Microdissection Objectives resolution
Leica Microdissection Objectives Laser microdissection has developed into one of the most important methods of specimen preparation. Specimens for gene expression profiling, proteomics, and various molecular diagnostic applications must consist of a homogenous cell population. For this purpose, selecting cells from tissue sections by means of microdissection has proven itself as a valuable tool. Increasingly, microdissection applications create molecular signatures as the basis for functional studies of clinical images or agent analysis. By using microdissection, a separate analysis of different cells from mixed tissues can provide a comprehensive view into tumor genesis. High quality optics are at the heart of laser microdissection, and image quality must meet the expected high standards of scientists. Also, high transmission in the UV range is required, coupled with excellent imaging of the UV laser through the objective. Leica Microsystems has developed a new set of objectives that optimize these functions and provide the best possible performance for single cell or larger tissue area microdissection. Leica Microsystems new SmartCut Series of Objectives range from 6.3x to 150x. Leica SmartCut objectives enhance all applications of proteomics including the selection of single chromosomes from metaphase spreads. 2
HCX PL FLUOTAR UVI 6.3x /0.13 Microdissection HCX PL FLUOTAR L 40x/0.60 CORR XT a b Pre-LMD Post-LMD Cap H&E stained left ventricular section shown above, prior to and following laser microdissection. The specimen is then shown in the cap. (a) Microdissection of a blood vessel; (b) Microdissection of a group of cardiac myocytes. The bar represents 100 µm. M r a IPG 3-10 NL x10 3 220 97 66 45 30 21.5 14.3 b IPG 3-10 NL Two-dimensional gel separations from H&E stained sections either (a) scraped or (b) laser microdissected. Protein spots were visualized by silver staining. 50 µm Section of muscle (human) following dual histochemistry for cytochrome c oxidase (COX) and succinate dehydrogenase (SDH), both mitochondrial enzymes. The blue muscle fibre is deficient in COX. Used with permission of Dr. G. Borthwick, Institute of Human Genetics, University of Newcastle Used with permission of Drs. A.I. De Souza and M.L. Rose. Images courtesy of Heart Science Centre, Harefield Hospital: De Souza, A.I.; McGregor, E.; Dunn, M. J.; Rose, M. L., Proteomics, 4, 578-86 (2004). Leica Microsystems UVI 6.3x objective was specifically designed for process applications in proteomics. High UV transmission at low magnification makes this objective a sophisticated tool for dissecting larger tissue areas from frozen or paraffin sections. High quality imaging of the laser focus in the specimen plane allows the user to effortlessly dissect thicker specimens without a significant widening of the dissection line. The clearly illuminated field of view also makes this objective an important tool for generating a good overall view of the cell(s) of interest. Leica Microsystems new 40x objective sets new standards in microdissection. Providing excellent quality and the highest UV transmission, this objective is the standard tool for almost all microdissection applications. Leica Microsystems 40x objective dissects even complex materials with a low dissecting line width. The 40x objective operates with a low effective laser aperture, which allows the laser beam to be precisely positioned in thicker specimens. This also makes this objective well-suited for laser ablation, among other techniques. The objective s correction ring ensures consistently excellent image quality. Order No. 11518145 Order No. 11506208 3
HCX PL FLUOTAR L 63x/0.70 CORR XT HCX PL FLUOTAR L 150x/0.90 Microdissection 150x HCX PL Fluotar, Metaphase Chromosomes on PET membrane slides Astrocytes: Immunfluorescence with GFAP primary and Cy3 secondary antibody Before cut During cut RT-PCR analysis of microdissected astrocytes from mouse cortex Cutting line on a 15 µm rat colon paraffin section stained with H&E C 1 2 C 1 2 C 1 2 Cyclophilin GFAP Neurofilament C: Whole brain control 1; 2: Astrocytes, microdissected (n= 10 cells) : no cdna input Small dissections down to a width of one micrometers are required to select cells in the range of 5-20 µm in diameter. Dissecting single cells of this width requires special optics. With Leica Microsystems 63x HCX PL FLUOTAR objective, these requirements are achieved with brilliant image quality. The large working distance of this objective allows easy and safe microdissection when working with groups or individual cells. Leica Microsystems HCX PL FLUOTAR 150x/0.9 Microdissection objective represents a special development. This150x magnification objective with high aperture creates dissecting line widths in the specimen of less than one micrometer. Leica designed the objective as a dry objective, which makes working at high magnification practical and easy. At the same time, this objective features an extremely brilliant image quality at an aperture of 0.9. Whether the tasks involve dissecting single astrocytes or chromosomes, this objective is the perfect solution whenever highest magnifications with extremely small dissecting widths are required. Order No. 11506222 Order No. 11506214 4
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