Animals were picked at the indicated age and transferred to a 24-well plate containing HS buffer (100 mM Tris-HCl, pH 7.4, 17 mM NaCl and 1% cholesterol supplemented with bacteria). Animals were then subjected to a 37°C HS for 6 h (or 9 h in the experiment depicted in Fig. S3 ). HS buffer was supplemented with SYTOX orange (Invitrogene) and animal survival was scored by monitoring dye uptake using a Leica M165 FC fluorescent stereoscope with a TXR filter. Fluorescent animals were scored as dead. >45 animals per experimental condition were scored.
M165 fc fluorescent
Manufactured by Leica camera
The M165 FC fluorescent is a high-performance microscope system designed for a wide range of fluorescence imaging applications. It features a powerful LED illumination system, providing bright and uniform illumination across the entire field of view. The system is equipped with a high-resolution digital camera and advanced imaging software, enabling users to capture detailed and accurate fluorescence images.
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Lab products found in correlation
4 protocols using m165 fc fluorescent
1
Heat Stress Assay for Nematode Survival
2
Imaging Fluorescent Transgenic Animals
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Age-synchronized animals expressing pwIs98(YP170::tdimer2) were fixed as in Karady et al. (2013 (link)) and imaged using a Leica M165 FC fluorescent stereoscope with a TXR filter. Pictures were analyzed using imageJ software (NIH).
3
Fluorescent Sexing of A. gambiae
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A. gambiae was fluorescently sorted, sexed, and imaged using the Leica M165FC fluorescent stereomicroscope using a Leica DMC2900 camera. Fluorescence was visualized using the CFP/YFP/mCherry triple filter and was sexed by examination of pupal genital terminalia. In cases where sex was indeterminable by pupal phenotype, genotype was validated by Y-chromosome PCR (see below).
4
Quantifying Plasmid Retention in RhiP Barley
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To assess plasmid loss in AcCherry (pSIR02) cells recovered from the RS and RA fractions of RhiP barley from flow-cytometry experiments, cell suspensions were plated on nonselective UMS agar and incubated 2 d. Sixty colonies recovered from each fraction of three RhiP plants were patched onto UMS agar supplemented with kanamycin, and onto nonselective UMS agar as a control. As the kanamycin resistance gene was expressed from pSIR02, resistant colonies were scored as having retained the plasmid. Silencing of pSIR02 was also assessed in the RS and RA fractions recovered from three wild-type and RhiP barley plants by plating cell suspensions on UMS agar supplemented with kanamycin or the same media additionally containing 1 μM SI. Plates were imaged using a Leica M165 FC fluorescent stereo microscope fitted with a CLS100x cold light source and DFC310 FX. To assess GFP induction, dual-channel images captured using dsRed and GFP filters were converted to 8-bit using ImageJ (55 (link)). mCherry+ colonies were demarcated for analysis in the dsRed channel using the thresholding tool (gray value > 20 and area > 100 pixel2) and these coordinates were superimposed over the GFP channel images. Colonies were scored as GFP+ if the maximum gray value exceeded 25. At least 32 colonies were analyzed per biological replicate. The raw data are provided (Dataset S6 ).
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