Whole mount immunohistochemistry of inner ear was performed as previously described [44] . Briefly, the inner ears were dissected in cold PBS buffer shortly after P30 mice were euthanized by C0 2 inhalation. Temporal bone was removed prior to overnight fixation in paraformaldehyde (4 % v/v in PBS) at 4 °C. The sensory epithelium was fine dissected, blocked, and permeabilized by incubation in blocking buffer (normal goat serum in 0.1% triton) for 2 hours at temperature. Samples were incubated with indicated primary antibody overnight at 4°C. After a brief wash in PBSX1, samples were incubated with secondary antibody for 2 hours at room temperature. To visualize F actin, this was followed by incubation for 1 hour at room temperature in phalloidin conjugated to Alexa Fluorophores (Life Technologies). The stained samples were mounted on Histobond microscope slides (Marienfeld GmbH) using Prolong Gold (Thermo Scientific) and dried overnight at room temperature. Image acquisition was performed with a confocal laser microscopy system (LSM800 Carl Zeiss).
Histobond microscope slides
Manufactured by Marienfeld
Sourced in Germany
Histobond microscope slides are a type of glass slide used for mounting and analyzing samples in microscopy. They provide a flat, smooth surface for specimens to be placed and observed under a microscope. The slides are designed to be compatible with a variety of microscopy techniques.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluorophore, by Thermo Fisher Scientific (2 mentions)
Lsm 800, by Zeiss (2 mentions)
Prolong gold, by Thermo Fisher Scientific (2 mentions)
Nis element, by Nikon (1 mentions)
Phosphate buffered saline (pbs), by Santa Cruz Biotechnology (1 mentions)
Maa lectin 1, by Vector Laboratories (1 mentions)
Vectashield antifade mounting medium with dapi, by Vector Laboratories (1 mentions)
5 protocols using histobond microscope slides
1
Whole Mount Immunohistochemistry of the Inner Ear
2
Whole Mount Immunohistochemistry of Inner Ear
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Whole mount immunohistochemistry of inner ear was performed as previously described (Dror et al., 2010 (link)). Briefly, the inner ears were dissected in cold PBS buffer shortly after P30 mice were euthanized by CO2 inhalation. Temporal bone was removed prior to overnight fixation in paraformaldehyde (4% v/v in PBS) at 4°C. The sensory epithelium was fine dissected, blocked, and permeabilized by incubation in blocking buffer (normal goat serum in 0.1% triton) for 2 h at room temperature. Samples were incubated with indicated primary antibody overnight at 4°C. After a brief wash in PBSX1, samples were incubated with secondary antibody for 2 h at room temperature. To visualize F actin, this was followed by incubation for 1 h at room temperature in phalloidin conjugated to Alexa Fluorophores (Life Technologies). The stained samples were mounted on Histobond microscope slides (Marienfeld GmbH) using Prolong Gold (Thermo Scientific) and dried overnight at room temperature. Image acquisition was performed with a confocal laser microscopy system (LSM800 Carl Zeiss).
3
Colorectal Cancer Sphere Formation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
SW480 cells (1 × 104) were seeded in 6-well ultralow attachment plates (SPL 3D™ Cell Floater, #39706; SPL Life Sciences, Gyeonggi-do, South Korea) and incubated with media supplemented with 10 % FBS. Sphere formation was observed after seven days, and sphere diameters were measured using NIS-Elements image analysis software (Nikon). For immunocytochemistry, 3D spheres were fixed using 3.7 % paraformaldehyde for 15 min, and spheroids were added to adhesive microscope slides (HistoBond® microscope slides, #0810001; MARIENFELD, Lauda-Königshofen, Germany), followed by immunocytochemistry.
4
Cryopreserved Organ Sectioning and Staining
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After harvesting, organs were fixed in 4% paraformaldehyde (PFA) solution in PBS (Santa Cruz Biotechnology, Inc., Heidelberg, Germany) for 1 day at 4 °C, immersed in a 30% (w/v) sucrose solution in PBS for 2 days at 4 °C, and frozen by vapor of liquid nitrogen, then stored at −80 °C until sectioning. Organs were embedded into Frozen Section Compound (Leica FSC 22 Clear, Leica Microsystems srl, Italy) and 20 µm slices were prepared using cryostat instrument. The sections were then placed on HistoBond® microscope slides (Marienfeld, Germany) and stained with Haematoxylin‐Eosin (Modified Gill's Type II Haematoxylin; Eosin G) or 4’,6‐diamidion‐2‐phenylindole (DAPI) and Luciferase antibody (Primary Antibody, Red Firefly Luciferase Polyclonal Antibody, eBioscience, 1:50; Secondary Antibody, Goat Anti‐Rabbit IgG Cross‐Adsorbed Alexa 594, Invitrogen, 1:1000) for tumor recognition. Slices were imaged using confocal microscopy (Nikon A1).
5
Lectin-Binding Assay for Cell Surface Glycans
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The lectin-binding assay was conducted as previously reported (12 (link)). For flow cytometry, cultured GAPDH-B4GALNT2 tagging and wild-type DF-1 cells were trypsinized and washed with 1 × PBS. Thereafter, cells were treated with green fluorescent Maackia Amurensis (MAA) Lectin I (20 μg/mL; Vector Laboratories FL-1311, Burlingame, CA, United States) for 20 min at 4°C, washed in 1 × PBS, and analyzed. For microscopy, cells were trypsinized, washed with 1 × PBS, placed on HistoBond microscope slides (Paul Marienfeld, Lauda-Konigshofen, Germany), and dried overnight at room temperature. After hydration, cells were treated with MAA-lectin I (20 μg/mL) for 1 h at room temperature, washed three times with 1 × PBS, mounted using VECTASHIELD Antifade Mounting Medium with DAPI (Vector Laboratories H-1200), and imaged under a fluorescence microscope.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!