A 1.5 glass coverslip was functionalized with 3-aminopropyltriethoxysilane (APTES; Sigma-Aldrich) by soaking in 10% APTES in ethanol for 5 min. The coverslip was then rinsed with deionized water and left to dry. The coverslip was then fixed onto an ibidi sticky-Slide VI 0.4 forming six microfluidic channels. First, 100 μl of phosphate-buffered saline (PBS) was pipetted through a channel. Next, the channel was filled with 50 μl of a solution (5 mg/liter) of nIRCats and left to incubate at room temperature for 5 min. The channel was rinsed using three 50 μl of PBS washes, keeping the channel filled with solution at all times. The surface-immobilized nIRCats in PBS were imaged on an epifluorescence microscope with an excitation of 721 nm and a Ninox VIS-SWIR 640 camera (Raptor). One end of the flow channel was connected to a syringe pump (Harvard Apparatus) using Luer lock fittings. Before the start of image acquisition, the opposite flow reservoir was filled with PBS, and the pump was set to refill mode at a volumetric flow rate of 40 μl min−1. Once the liquid in the reservoir was depleted, 40 μl of 10 μM DA in PBS was added. The process was repeated using alternating additions of 80 μl of PBS washes and 40 μl of DA solution.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.