After the imaging procedure, mice were perfusion fixed using 4% paraformaldehyde (PFA, Thermo Fisher, Loughborough, UK) in PBS. After enucleation, the cornea and lens were removed under direct visualisation with an operating microscope in 4% PFA in PBS. After fixation overnight, the eyecups were cryoprotected using a 10-30% sucrose gradient. Eyecups were embedded in optimal cutting temperature (OCT) compound (Tissue-Tek, Sakura Finetek, The Netherlands), frozen on dry ice and stored at -80°C until sectioning. Eyecups were cryosectioned into 16μm sections and affixed to poly-L-lysine coated glass slides (Polysine®; Thermo Scientific, Loughborough, UK). The sections were air-dried and then stored at -20°C until further histological processing.
Polysine
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
Polysine is a laboratory product designed for the adhesion of biological samples to microscope slides. It provides a positively charged surface that enhances the attachment of tissues, cells, and other specimens during various analytical and diagnostic procedures.
Automatically generated - may contain errors
Lab products found in correlation
Paraformaldehyde (pfa), by Thermo Fisher Scientific (2 mentions)
Tissue tek, by Sakura Finetek (2 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Lsm 510 meta, by Zeiss (2 mentions)
Poly l lysine coated coverslips, by BD (1 mentions)
Ellipse microscope, by Nikon (1 mentions)
Vectashield mounting medium, by Thermo Fisher Scientific (1 mentions)
Vectashield mounting medium, by Vector Laboratories (1 mentions)
Glass coverslips, by Electron Microscopy Sciences (1 mentions)
Poly d lysine pdl, by Merck Group (1 mentions)
Cell tak, by Corning (1 mentions)
6 protocols using polysine
1
Fixation and Sectioning of Mouse Eyes
2
Fixation and Sectioning of Mouse Eyes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
After the imaging procedure, mice were perfusion fixed using 4% paraformaldehyde (PFA, Thermo Fisher, Loughborough, UK) in PBS. After enucleation, the cornea and lens were removed under direct visualisation with an operating microscope in 4% PFA in PBS. After fixation overnight, the eyecups were cryoprotected using a 10-30% sucrose gradient. Eyecups were embedded in optimal cutting temperature (OCT) compound (Tissue-Tek, Sakura Finetek, The Netherlands), frozen on dry ice and stored at -80°C until sectioning. Eyecups were cryosectioned into 16μm sections and affixed to poly-L-lysine coated glass slides (Polysine®; Thermo Scientific, Loughborough, UK). The sections were air-dried and then stored at -20°C until further histological processing.
3
Quantification of Micronuclei and Chromatin Bridges
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For the quantification of micronuclei, cells were treated with 2 μg ml−1 Cytochalasin B on poly-L -lysine-coated coverslips (BD Bioscience) for 12 h before being fixed and mounted with Vectashield mounting medium with DAPI (Vector laboratories). Micronuclei in binucleated cells were scored using the micronucleus assay21 (link). For the quantification of chromatin bridges and binucleation, cells were harvested following asynchronous growth incubated on Polysine (Thermo Scientific) slides, fixed and mounted with Vectashield mounting medium with DAPI. Slides were scored using a Nikon Ellipse Microscope. Binucleation was defined and scored as in the micronucleus assay.
4
Fluorescence Visualization of Antimicrobial Peptide Action
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The fluorescence experiments were performed with slight modification as described previously [22 (link)]. Briefly, the bacteria (107 CFU/mL) were incubated with 0.5 ×MIC antimicrobial peptides for 30 min and then treated with BODIPY-labeled vancomycin (2 μg/mL) for 30 min. Samples treated with BODIPY-labeled vancomycin only served as a control. After incubation, bacteria were centrifuged at 8000× g for 5 min, washed three times with PBS to remove unbinding fluorescence dye, and resuspended by PBS. The resuspended solution were loaded on the glass slides (Polysine™, Thermo Fisher Scientific, Waltham, MA, USA) and visualized under confocal laser scanning microscope (LSM 510 META, Carl Zeiss, Jena, Thüringen, Germany) equipped with a 64× oil objective lens (Carl Zeiss, Jena, Thüringen, Germany).
5
Decellularized Bovine Achilles Tendon Sections
6
Immobilizing Bdellovibrio bacteriovorus for AFM
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Glass slides were washed,
rinsed with ethanol, and plasma-cleaned. They were then coated with
the polyphenolic adhesive Cell-tak (Corning) according to the manufacturer’s
directions. Cleared predatory B. bacteriovorus prey lysate was centrifuged at 3800 × g for
15 min to remove the remaining E. coli. The resulting supernatant was centrifuged for another 15 min at
5400 × g to pellet the predatory cells. These
cells were resuspended in HM at one fourth their original volume,
and the concentrated suspension was placed on the Cell-tak-coated
slide for 40 min. HI B. bacteriovorus was centrifuged at 5400 × g for 4 min, and
the pellet was resuspended in HM and then placed on Cell-tak-coated
slides for 2 min. Slides for both predatory and HI B. bacteriovorus were rinsed with deionized water
and immediately rewetted with fresh HM buffer for AFM analysis.
HI cells in LB buffer were also added directly to poly-l -lysine-coated slides (Polysine, Thermo Scientific). This process
was completed in the same manner as cellular adhesion to the Cell-tak
slides, and no difference in the extent of cell adhesion nor the shape
of force curves was seen between the two adhesives.
rinsed with ethanol, and plasma-cleaned. They were then coated with
the polyphenolic adhesive Cell-tak (Corning) according to the manufacturer’s
directions. Cleared predatory B. bacteriovorus prey lysate was centrifuged at 3800 × g for
15 min to remove the remaining E. coli. The resulting supernatant was centrifuged for another 15 min at
5400 × g to pellet the predatory cells. These
cells were resuspended in HM at one fourth their original volume,
and the concentrated suspension was placed on the Cell-tak-coated
slide for 40 min. HI B. bacteriovorus was centrifuged at 5400 × g for 4 min, and
the pellet was resuspended in HM and then placed on Cell-tak-coated
slides for 2 min. Slides for both predatory and HI B. bacteriovorus were rinsed with deionized water
and immediately rewetted with fresh HM buffer for AFM analysis.
HI cells in LB buffer were also added directly to poly-
was completed in the same manner as cellular adhesion to the Cell-tak
slides, and no difference in the extent of cell adhesion nor the shape
of force curves was seen between the two adhesives.
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!