Assays were performed as previously described (Hrycaj et al., 2018a (link)). Non-tissue culture treated polystyrene 96-well flat bottom microtiter plates (Denville, # T1097) were coated and incubated with bovine plasma fibronectin (Sigma-Aldrich, #F1141) at 20 μg/ml for 1 h at 37°C. Plates were then blocked with 100 μl/well of 1% BSA (Sigma-Aldrich, #A7906) in serum-free DMEM/F12 (Gibco, #11320033) for 30 min at 37°C. Fibroblasts were seeded at 10,000 cells per individual well. Plates were centrifuged (top side up) at 10 × g for 5 min to reduce the variability inherent in the settling of cells onto the plate surface and were incubated for 1 h at 37°C with 5% CO2. Non-adherent cells were removed by centrifugation (top side down) at 48 × g for 5 min. Adherent cells remaining on the plate were fixed and stained with 1% formaldehyde (Fisher Sci., #BP228-100), 0.5% crystal violet (Sigma-Aldrich, #C6158), 20% MeOH followed by PBS washes. Individual wells were imaged on a Leica MZ125 dissecting microscope and manually quantified using ImageJ 2.0 cell counter function.
Mz12.5 dissecting microscope
Manufactured by Leica
Sourced in Germany
The Leica MZ12.5 is a dissecting microscope designed for precise observation and examination of samples. It features a maximum magnification of 125x and a working distance of up to 114mm, allowing for detailed analysis of a wide range of specimens. The microscope is equipped with LED illumination for optimal visibility and clarity.
Automatically generated - may contain errors
Lab products found in correlation
C6158, by Merck Group (1 mentions)
F1141, by Merck Group (1 mentions)
A7906, by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Mp e 65 mm macro lens, by Canon (1 mentions)
5d mark 3, by Canon (1 mentions)
Illustrator cs6, by Adobe (1 mentions)
Eos 550d, by Canon (1 mentions)
4 protocols using mz12.5 dissecting microscope
1
Quantifying Fibroblast Adhesion on Fibronectin
2
Assessing Wnt Signaling in Zebrafish
3
Whole-mount Hoxa5 embryo imaging
4
Multidimensional Imaging Techniques for Specimen Documentation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Handmade draft drawings were produced using a LEICA MZ12.5 dissecting microscope and illustrated with the aid of a drawing tube (Leica, Wetzlar, Germany). Photographs were taken using Canon EOS 550D or 5D Mark III digital cameras (Canon, Tokyo, Japan), coupled to a Canon 50 mm macro lens, a 100 mm macro lens, or a Canon MP-E 65 mm macro lens, all equipped with polarizing filters. Each specimen was photographed under dry condition and covered with a thin film of ethanol. When available, both imprints were photographed. These photographs were optimized using Adobe Photoshop CC 2015.5 (Adobe Systems, San Jose, CA, USA) and assembled, together with handmade drawings, into a single, multi-layered document. Reproduced photographs referred to as 'composites' are a combination of photographs of a dry specimen and the same under ethanol. In addition to traditional photographs, we computed Reflectance Transforming Imaging (RTI) files for details of several specimens. This corpus of data was used to produce illustrations using Adobe Illustrator CS6 (Adobe Systems, San Jose, CA, USA). Multi-layered documents (photographs only) and RTI files are provided in the associated Dryad dataset (58) . Investigated specimens are listed in the section 2.1.2.
Measurements were based on complete specimens illustrated herein and are provided in the following format: minimum/average/maximum.
Measurements were based on complete specimens illustrated herein and are provided in the following format: minimum/average/maximum.
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