For platelet adhesion assays, video microscopy and immunofluorescence microscopy experiments, #1.5 glass coverslips (Warner Instruments) or #1.5 Glass Bottom Dishes (MatTek) were coated with 50 μg/ml human fibrinogen (FIB3, Enzyme Research Labs) or 100 μg/ml fibrillar collagen (Chronolog) and blocked with fatty-acid free BSA (Sigma) prior to platelet seeding, as described [18 (link)]. Platelet static adhesion, immunofluorescence and live video microscopy were performed using Kohler-illuminated Nomarski differential interference contrast (DIC) optics with a Zeiss 63× oil immersion 1.40 NA plan-apochromat lens on a Zeiss Axiovert 200M microscope as previously described [19 (link)-21 (link)]. The surface areas of individual platelets were measured using Image J software and plotted as previously described [18 (link)]. For acK immunofluorescence experiments, fixed platelets were blocked, permeablized and stained in blocking buffer (PBS + 0.1% SDS + 1% BM). Super resolution-structured illumination microscopy (SR-SIM) experiments were carried out on a Zeiss Elyra SR-SIM microscope as previously described [17 (link), 18 (link)].
Protocol detail
Platelet Adhesion Assays and Microscopy
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Assays
Buffer
Dishes
Enzyme
Fibrillar collagen
Fibrinogen
Free fatty acid
Human
Illumination microscopy
Immersion
Immunofluorescence
Immunofluorescence microscopy
Lens
Microscope
Optics
Platelet adhesion
Platelets
Video microscopy
Corresponding Organization : Cardiovascular Institute of the South
Other organizations : Université de Lille, Oregon Health & Science University
Top 5 similar protocols
Variable analysis
independent variables
- Coating of #1.5 glass coverslips or #1.5 Glass Bottom Dishes with 50 μg/ml human fibrinogen or 100 μg/ml fibrillar collagen
- Platelet static adhesion
- Immunofluorescence
- Live video microscopy
- Surface areas of individual platelets measured using Image J software
- Blocking of coated surfaces with fatty-acid free BSA prior to platelet seeding
- Use of Kohler-illuminated Nomarski differential interference contrast (DIC) optics with a Zeiss 63× oil immersion 1.40 NA plan-apochromat lens on a Zeiss Axiovert 200M microscope
- Fixation, permeabilization, and blocking of platelets for immunofluorescence experiments
- Use of Super resolution-structured illumination microscopy (SR-SIM) for specific experiments
- Positive control: Not explicitly mentioned
- Negative control: Not explicitly mentioned
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!