DRGs (L5 for SNL and lumbar for STZ-induced DM) of the experimental animals were embedded in paraffin, sectioned for 5 µM sections and probed after deparaffinization with isolectin Griffonia simplicifolia (IB4) conjugated with Alexa 488 (1:200; Vector Laboratories, USA) and antibodies against CGRP (1:10 000, Peninsula Laboratory, USA). To visualize bound CGRP primary antibodies, slides were incubated with biotinylated Goat Anti-Rabbit IgG Antibody (Vector Laboratories, USA) and ABC-peroxidase conjugate (VECTASTAIN Elite ABC-Peroxidase Kit antibody, Vector Laboratories, USA) followed by chromogenic visualization with DAB staining kit (Vector Laboratories, USA) according to the manufacturer’s instructions. In the slides labeled with IB4-Alexa488, nuclei were stained with DAPI. The slides were mounted with Immu-Mount (Thermo Scientific, USA) or Coverquick 2000 (VWR, Finland) and imaged with fluorescent (Zeiss Imager M2 Axio, Carl Zeiss, Germany) or bright-field (Olympus BX-61, Olympus, Japan) microscopes. The number of marker-positive neurons in a section of a DRG was calculated manually and normalized to the total number of sensory neurons in this section. The results of the analysis of two to four sections per ganglion were averaged and used in further calculations.
Coverquick 2000
Manufactured by Avantor
Sourced in United States, Germany
The Coverquick 2000 is a lab equipment product that serves as an automated slide coverslipping device. It is designed to efficiently and consistently apply coverslips to prepared microscope slides.
Automatically generated - may contain errors
Lab products found in correlation
Biotinylated goat anti rabbit igg antibody, by Vector Laboratories (1 mentions)
Axio imager m2, by Zeiss (1 mentions)
Dab staining kit, by Vector Laboratories (1 mentions)
Alexa 488, by Vector Laboratories (1 mentions)
Immu mount, by Thermo Fisher Scientific (1 mentions)
Hemacolor, by Merck Group (1 mentions)
2 protocols using coverquick 2000
1
Histological Analysis of Neuronal Markers
2
Centrifugation of Fluid Samples
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The second specimen was placed in a modified commercial salad spinner (26 cm diameter, Zyliss cat: 15201; Diethelm Keller brands, Zurich, Switzerland) (Figure 1). This kitchen device is accelerated by pulling a handle continuously to half distance 90-100 times per min (controlled by a wall watch). On average, the basket rotated at 127g (1150 rpm) for 5 min as measured by a digital tachometer (DT-2234C; Rinch Industrial, Shanghai, China, accuracy 1 rpm). The same duration and rotation were followed for all fluid samples (including CSF). Styrofoam cushions of 5, 9, 3, 9, 2 cm were fitted to the basket with rubber bands (Figure 2), which also held the cell concentrators in place. All slides were stained with a commercial Romanowsky-type stain (Hemacolor; Merck, Darmstad, Germany) and mounted with mounting media (Cover-quick 2000; VWR Chemicals, Fontenay-sous-bois, France).
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