Scroll down the linked page to find the version appropriate to your operating system. LAS X Core: Free software from Leica to open images acquired on the Leica SP8 confocal or any microscope controlled by LAS X. ZEN Lite : Free software from Zeiss to open. Imaris Viewer: A free version of Imaris with limited capabilities useful for quickly viewing Imaris files or raw data. Ilastik: Free software that uses machine learning for interactive object recognition and segmentation. It's absolutely necessary for playing videos created using our Core's version of Arivis Vision4D at the Lightsheet workstation.ĬellProfiler: Free, open-source software from the Broad Institute for segmentation and quantitative analysis of microscope image data. VLC: VLC media player is a free and cross-platform multimedia player that plays most multimedia files as well as DVDs, Audio CDs, VCDs, and various streaming protocols. Recommended for all CDB Micro Core users. Includes utilities for 3D viewing, video editing, measuring colocalization among others. Very handy for new users who don't want to download individual plugins. Finally don’t forget to use the proper longpass or bandpass filter to reject your fluorescence excitation source.Fiji: ImageJ plus a large bundle of plugins, all packaged together in one easy-to-download application. Depending on the turbidity of your liquid media, you should control the thickness by selecting the best demountable cuvette because the scattering coefficients in the UV in turbid media are relatively high - meaning UV excitation is attenuated within a few tens to hundred micron of thicknesses - and the excess thickness becomes redundant. Since you are looking at fluorescence, I assume you are using UV excitation. Also you may think about sonicating your lipid-water mixture to produce more repeatable droplet sizes by using different frequencies. You can get demountable curettes with different cross-sectional thicknesses and therefore allowing you to trap a constant thickness of liquid mixture for your experiment. You should control the thickness of your liquid mixture with demountable cuvettes ( ). Also your approach of plating a microcope slide with or without a cover slip gave you very unpredictable liquid thicknesses it’s uncontrolled and not easily repeatable. You didn’t mention the size range of the lipid droplets you are investigating. If anyone has any suggestions for a potential solution or an alternative method to create a triglyceride calibration curve using Nile Red and fluorescence microscopy it would be greatly appreciated! Thank you in advance! The tones, in particular, are sublime and I absolutely love the look of the files this camera produces. Essentially, I am looking for any ideas regarding a possible solvent (suspension media) that would allow the stained lipid droplets to remain suspended as spheres in the solution and not rise (or rise very slowly) to the surface of the liquid, causing them to lose their shape. With such great glass on the front of the Leica Q, a 24MP CMOS sensor and a system optimised for the 28mm lens, the image quality is stunning. When the images were attempted without a coverslip the droplets moved too much (or stuck to the sides of the slide well) to obtain good quality images for quantification. When we add a small volume of triglyceride to water, with the intent to simulate the “lipid droplets” found in the animals, plate the solution on a microscope slide and place a cover slip on top, the droplets naturally rise to the surface of the water and flatten out against the cover slip making it difficult to estimate their volume. In order to make the calibration curve we need to produce Nile Red stained spherical lipid droplets of various sizes. I am working on creating a calibration curve that relates the volume of Nile Red stained triglyceride droplets, in various tissues of small invertebrates, to a fluorescence intensity using a Leica microscope with a fluorescence detector.
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