The UCIM microscopy facility offers researchers a wide variety of optical microscopes covering most of the techniques used in biomedical research. From basic techniques such as brightfield or reflected light microscopy to the most advanced ones such as FRET, confocal fluorescence microscopy, calcium probes, live cell observation, etc. <\/p>\n
The aim of the service is to provide comprehensive support to researchers and make it possible to incorporate microscopy into their projects, advising and answering any questions they may have in this field. From sample preparation to image analysis.<\/p>\n<\/div>
The preferred mode of use of the equipment is self-service, once the necessary training has been received. Alternatively, there is the possibility of sessions assisted by the section’s technicians.
\nThe equipment can be booked through the section’s web application.
\nTo register for the booking application, you must send an e-mail to microscopia.ucim@uv.es in accordance with the general rules of use.<\/p>\n<\/div>
– Bright-field microscope<\/strong> – Confocal microscope SP8<\/strong> – DMi8 inverted live cell microscope – calcium probes<\/strong> – Fluorescence inverted microscope<\/strong> – Sample preparation: cryostat, microtome and paraffin embedding system<\/strong> Leica SP8 Confocal Microscope<\/a> DMi8 microscope Leica LAS-X<\/a><\/strong>: Fiji<\/a><\/strong>: Ilastik<\/a><\/strong>:
\nThe bright-field microscope allows us to view stained or naturally pigmented samples that are highly contrasted. The source of illumination is white light. The components of the sample are visualised thanks to the differences in contrast between them and the surrounding medium.<\/p>\n
\nThe confocal microscope allows us to visualize images with different fluorescent labelling, therefore obtaining images of great sharpness and quality because the pictures earned are not contaminated by light emitted outside the focal plane. Thanks to this confocal characteristic, we can perform three-dimensional reconstructions from optical sections.<\/p>\n
\nThe variety of fluorescent proteins and multicolour probes that have been developed allows virtually any molecule to be tagged. The ability to visualise protein dynamics in vesicles, organelles, cells and tissues has provided new information about the functioning of cells in both healthy and diseased situations. This information includes the spatio-temporal dynamics of processes, such as mitosis; studies on the dynamics of ions, such as Ca+\u2082 and changes in the cytoskeleton.<\/p>\n
\nFluorescence is one of the most widely used physical phenomena in biological and analytical microscopy, mainly because of its high degree of sensitivity and specificity. Fluorescence microscopy allows users to determine the distribution of a single molecule, its quantity and its location within a cell.
\nFluorescence microscopes used in research applications are based on a series of optical filters. The filters are usually inserted in a filter block. While the excitation filter selects the wavelengths that excite a particular fluorophore within the sample, the emission filter acts as a kind of quality control, as it only allows the wavelengths of interest emitted by the fluorophore to pass through.<\/p>\n
\nThe cryostat, also known as cryotome, is a freezing microtome used for cutting slices of frozen material. It\u2019s located on a cold room\u2019s interior.
\nThe microtome is used to make histological cuts that vary in thickness. It\u2019s a precision tool allowing to obtain uninterrupted series of cuts of even and specific thickness.
\nThe paraffin embedding system is a perfusion of paraffin itself within tissues which creates a homogeneous medium allowing to make cuts of little thickness with ease and precision.<\/p>\n<\/div>Tutorials and Resources<\/h5>\n
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\nInverted microscope DMi8 per living cell<\/a>
\nLeica DMI 3000-B inverted fluorescence microscope<\/a><\/p>\n\n
\nBasic configuration of LAS-X software for image acquisition<\/a><\/p>\n\n
\nProgram that, with its free version, allows viewing, exporting and editing images in Leica format.<\/p>\n
\nIamgeJ2 distribution with high quality plugins installed to facilitate image analysis.<\/p>\n
\nProgram that uses machine learning algorithms to easily segment, classify, track, and count cells and other experimental data.<\/p>\n