NTA was performed using a Nanosight LM10 instrument equipped with a 405 nm laser and a Hamamatsu C11440 ORCA-Flash 2.8 camera (Hamamatsu) with Nanoparticle Tracking Analysis (NTA, Malvern Instruments, UK) and data was analyzed with the NTA software 2.3 following the manufacter’s instructions. To define the size and concentration of the particles, the samples were diluted appropriately with Milli-Q water (Milli-Q Synthesis, Merck Millipore, Massachusetts, USA) to give counts in the linear range of the instrument. The particles in the laser beam undergo Brownian motion, and a video was recorded for 60 s in triplicate.
C11440 orca flash 2.8 camera
Manufactured by Hamamatsu Photonics
Sourced in United Kingdom
The C11440 ORCA-Flash 2.8 is a scientific-grade CMOS camera that offers a sensor with 2.8 megapixels. It features a large active area, high quantum efficiency, and low read noise, providing high-quality image capture for a variety of applications.
Automatically generated - may contain errors
5 protocols using c11440 orca flash 2.8 camera
1
Nanoparticle Tracking Analysis Protocol
2
Quantitative Analysis of Extracellular Vesicles
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The size and number of EVs were determined using a Nanosight LM10 instrument equipped with a 405 nm laser and a Hamamatsu C11440 ORCA-Flash 2.8 camera (Hamamatsu) with Nanoparticle Tracking Analysis (NTA, Malvern Instruments, UK). Each sample was diluted appropriately with Milli-Q water (Milli-Q Synthesis, Merck Millipore, MA, USA) to give counts in the linear range of the instrument. The particles in the laser beam underwent Brownian motion, and a video was recorded for 60 s in triplicate. The analysis was performed by following the manufacturer’s instructions, and data were analyzed using version 2.3 of the NTA software.
3
Nanoparticle Size and Concentration via NTA
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NTA was performed using a NanoSight LM10 system (Malvern Instruments) equipped with a 405 nm laser and a Hamamatsu C11440 ORCA-Flash2.8 camera (Hamamatsu). Data was analyzed with the NTA software 2.3. Size and concentration of particles were determined by the following settings: camera level and detection threshold were set to maximum (15 or 16) and minimum (3–5), respectively; camera gain was set to 512; blur, minimum track length, and minimum expected size were set to “auto.” Readings were taken in triplicates during 60 s at 18.87 frames/s, at room temperature ranging from 23–25 °C.
4
Nanoparticle Tracking Analysis of EVs
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Size and number of EVs was determined using a Nanosight LM10 instrument equipped with a 405 nm laser and a Hamamatsu C11440 ORCA-Flash 2.8 camera (Hamamatsu) with Nanoparticle Tracking Analysis (NTA, Malvern Instruments, UK). Each sample was diluted appropriately with Milli-Q water (Milli-Q Synthesis, Merck Millipore, Massachusetts, USA) to give counts in the linear range of the instrument. The particles in the laser beam undergo Brownian motion, and a video was recorded for 60 s in triplicate. Analysis was performed following manufacturer’s instructions and data were analyzed using the version 2.3 of the NTA-software.
5
Immunofluorescent Staining of hESCs
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After fixation with 4% paraformaldehyde (PFA), hESC cells were washed and incubated for 1 h in a blocking solution composed of 10% donkey serum and 0.1% Triton-X100 in TBS. Cells were then incubated at 4 1C over night with the mouse anti SSEA4-PE conjugate (Life Technologies A14766, 1:200) and mouse anti Oct3/4 (Millipore MAB4401, 1:200) in combination with the donkey anti mouse AF488 (Life Technologies, 21202, 1:200) secondary antibody. DNA was then stained with DAPI and cultures were imaged using an inverted microscope (Olympus IX73 equipped with a Hamamatsu C11440 Orcaflash 2.8 camera).
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