The focus of our investigation was on thin-caliber cutaneous innervation implicated in pain (nociceptors) and on affiliated epidermal keratinocytes which have been shown to modulate the sensitivity of this innervation (Baumbauer et al., 2015 , Moehring et al., 2018 ). For these evaluations, complete high-resolution epifluorescence digital montages of immunolabeled biopsy sections were captured using a computer linked Olympus BX51-WI microscope equipped with conventional fluorescence filters, a Hamamatsu ER DVC high-speed camera, and 3-axis motorized stage system interfaced with Neurolucida software (MBF Bioscience, Essex, VT). The data collection and CMA platform were performed under completely blinded conditions.
Bx51 wi microscope
Manufactured by Hamamatsu Photonics
The BX51-WI is an upright microscope designed for a wide range of research applications. It features a sturdy, ergonomic design and high-quality optics for precise observation and imaging. The microscope is capable of various observation techniques, including bright-field, dark-field, and phase-contrast microscopy.
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2 protocols using bx51 wi microscope
1
Investigating Cutaneous Nociceptor Innervation
2
Quantifying Tumor Hypoxia Levels
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Size matched untreated tumours or tumours treated with TH-302 (50 mg/kg, i.p.) for 2 h were snap frozen in liquid nitrogen after injection of pimonidazole hypoxia marker (60 mg/kg, i.p., Bioconnect) 1 h prior to tumour excision. Tumour sections (7 mm) were fixed with acetone (4 °C), rehydrated in phosphate buffered saline (PBS), incubated with normal goat serum followed by incubation with the primary rabbit anti-pimonidazole antibody (1:250; HP3-1000, Bio-connect) overnight at 4 °C and secondary goat antirabbit Alexa594 (1:500, Invitrogen) for 1 h at room temperature. Sections were mounted using fluorescent mounting medium (DakoCytomation). Whole tumour cross-sections were scanned using an Olympus BX51WI microscope equipped with a Hamamatsu EM-CCD C9100 digital camera, a motorized stage (Ludl Mac 2000) and 10-fold objective. After scanning, sections were stained with haematoxylin and eosin for the identification of the viable tumour compartment by morphological criteria. Pimonidazole hypoxic fraction, i.e. percent tumour area stained with pimonidazole was defined as described previously in detail [34] .
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