Images were taken using a Leica MZ FLII stereomicroscope and the Leica Application Suite V3 software. Contrast and image size were adjusted with Adobe Photoshop CS5.1. All bar plots were generated using GraphPad Prism 7. All figures were generated with Adobe Photoshop or Adobe Illustrator.
Mzfl 2 stereomicroscope
Manufactured by Leica
The MZFL II stereomicroscope is a versatile laboratory equipment designed for magnified observation and examination of samples. It features binocular eyepieces and a range of objective lenses to provide variable magnification. The MZFL II is a tool for detailed visual analysis in scientific and research settings.
Automatically generated - may contain errors
4 protocols using mzfl 2 stereomicroscope
1
Stereomicroscopy Image Capture and Visualization
2
Transgenic Root Generation in D. trinervis
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The pDISC-ProDtAUX1:GUS and the DR5:Venus binary vectors were introduced into A. rhizogenes ARqua1 strain by electroporation. Transformation of D. trinervis was performed using the ex vitro method described in Imanishi et al. (2011 (link)). Transgenic roots were identified by detecting DsRED or Venus fluorescence using a MZFLII stereomicroscope (Leica) with GFP2 or G filter sets or using a blue LED bulb (Orbitec) and 2846 protection glasses with orange lens (3M).
3
Tumor Tissue Harvesting and Preservation
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When reaching endpoints, mice were sedated with CO2 inhalation followed by intracardiac perfusion with cold HBSS 1X. After harvesting the brain, the GFP+ tumor was cut into two parts, under the MZFL II stereomicroscope (Leica). The anterior part of the GFP+ tumor and the GFP− parenchyma were chopped and stored in TRI-reagent (Molecular Center Research, #TR 118) at −80 °C or directly snap-frozen in liquid nitrogen for RNA isolation. The posterior part was snap-frozen in dry ice-cooled-isopentane for histological studies. Brains were cryosectioned at a 12-µm thickness (Leica cryostat).
4
Dissociation and Sorting of GFP+ Tumor Cells
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After brain harvest, GFP+ tumors were dissected under a Leica MZFL II stereomicroscope. Tumor pieces were chopped and incubated for 5 min at 37 °C in an HBSS-papain-based lysis buffer (Worthington PAP) containing DNAse (0.01%, Worthington #LS002139) and L-Cystein (124 µg/mL, Sigma #C78805). Papain digestion was inhibited by ovomucoid (7 mg/mL, Worthington #LS003085). Tissue was further dissociated mechanically and centrifuged 300 × g, 10 min at 4 °C. Cells were resuspended in cold HBSS, a debris removal step was performed (Miltenyi #130-109-398) and blood cells were removed using a blood lysis buffer (Roche 11814 389001). After centrifugation, cells were resuspended in cold HBSS and incubated with the eBiosciences Fixable Viability Dye Fluor 450 or 660 (Invitrogen 65-0863), to label dead cells, and washed. Cells were then sorted using the MoFlo Astrios cell sorter (Beckman Coulter) or the S3e cell sorter (Biorad). Live cells were collected in HBSS 0.1% BSA precoated tubes, centrifuged, and resuspended in HBSS–0.1% BSA at a concentration of 1200 cells/µL. GFP+ and GFP− cells were collected separately for the scRNAseq ddSeq experiment.
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