Tumours were harvested on day 5 after ACT and fixed in 4% paraformaldehyde for 24 h, then dehydrated in 20% sucrose before embedding in optimal cutting temperature freezing media (Sakura Finetek). Next, 6 µm sections were cut on a CM305S cryostat (Leica), adhered to Superfrost Plus slides (VWR) and stored at −20 °C until further use. When thawed, slides were either fixed with ice-cold acetone and stained with rat anti-mouse I-A/I-E (1:50) and anti-rat IgG-Alexa Fluor 594 (1:100) or directly mounted with Vectashield Antifade Mounting Medium (Vector Laboratories). Images were acquired on an Axio Imager.M2 with a Colibri 7 LED illumination system (Zeiss) and analysed with ImageJ v.1.52i (http://imageJ.nij.gov/ij ).
Colibri 7 led illumination system
Manufactured by Zeiss
Sourced in Canada, Germany
The Colibri 7 LED illumination system is a compact and efficient light source designed for use in a variety of microscopy applications. It features a high-performance LED that provides stable and uniform illumination across a wide range of wavelengths. The system is designed for easy integration with microscopes and other laboratory equipment, offering a convenient and versatile solution for researchers and laboratory professionals.
Automatically generated - may contain errors
Lab products found in correlation
Optimal cutting temperature freezing media, by Sakura Finetek (1 mentions)
Cm305s cryostat, by Leica (1 mentions)
Superfrost plus slides, by Avantor (1 mentions)
Vectashield antifade mounting medium, by Vector Laboratories (1 mentions)
Axiocam 702 mono, by Zeiss (1 mentions)
Observer z1 inverted microscope, by Zeiss (1 mentions)
Axiocam 506 camera, by Zeiss (1 mentions)
3 protocols using colibri 7 led illumination system
1
Cryosectioning and Immunostaining of Tumours
2
Quantifying Angiogenesis in HUVEC Cultures
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The organization of HUVECs into capillary structures at the surface of individual cell sheets was evaluated daily for six days after seeding GFP-transduced HUVECs. Stitched images of the entire area of the tissues were taken using a Zeiss fluorescence microscope equipped with a Colibri 7 LED illumination system (Zeiss, Toronto, ON, Canada). Central regions of interest (ROI) (25 mm2) of the stitched pictures were then analyzed using the Angiogenesis Analyzer plug-in of ImageJ® software (NIH, Bethesda, MD, USA), as previously described [39 (link)]. GFP-positive structures such as master segments, length of the master segments, meshes, and master junctions were analyzed for each condition (n = 3 tissues/condition per time point).
3
Comprehensive Imaging of Biological Samples
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Images were acquired by scanning 36 tiles at 20× magnification, corresponding to a total area of 6.4 mm 2 (Fig. 1a), over the same spatial area of all serial sections of the tested antibody. Brightfield and fluorescence images were acquired using the Zeiss Observer.Z1 inverted microscope (Carl Zeiss, Oberkochen, Germany) equipped with the Colibri 7 LED illumination system (Carl Zeiss). For brightfield images, the Axiocam 506 camera (Carl Zeiss) was used, and fluorescence images were taken with the Axiocam 702 mono (Carl Zeiss) equipped with an excitation and emission filter set for the visualization of DAPI, FITC, Cy3, and Cy5. The 20× objective (LD Plan-Neofluar 20×/0.40 corr., D = 0-1.5 mm; Carl Zeiss), and the ZEN 3 blue software (Version 3.0; Carl Zeiss) was used for capturing both the brightfield and fluorescence images.
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