Al 1000 syringe pump

Fluo-3-loaded Meg-01 cells attached to slides were treated with 2.5 μm GsMTx-4 for 1 min or vehicle (HBSS) as necessary before inserting the slides into the parallel-plate flow chamber. To apply fluid shear stress, the reservoir was filled with either HBSS, Ca²⁺-free HBSS containing 1 mm EGTA or HBSS containing 2.5 μm GsMTx-4 as appropriate, and the AL-1000 syringe pump (World Precision Instruments) was set to draw fluid through the system at the shear rates of 1002.6, 2282.67, and 3989.248 s⁻¹, which represent shear conditions in normal arteries, moderately stenotic arteries, and severely stenotic arteries, respectively (56 ). The fluorescent signals were background-corrected, and fluorescence levels (F) were normalized against prestimulus fluorescence level (F₀) to yield F/F₀ values.

The optical design of the device is based on a simplified variant of the SPIM-Fluid imaging flow cytometer [20 (link)] and described in detail in our previous publication [14 (link)]. The major features are shown in Figure 1. We are making a simplified lightsheet microscopy system where the light sheet is formed by a pair of 50 mm cylindrical lens and illumination objective lens 4×/NA 0.13 Nikon objective. We used 488 nm laser Cobolt 06-MLD (HÜBNER Photonics, Kassel, Germany) with a 3.5× beam expander as illumination source. The detection system in this variant is based on 10×/NA 0.3 Nikon objective lens, a set of fluorescent filters, where MF530-43 (Thorlabs, Newton, NJ, USA) FITC line filter was used in our experiments, a tube lens with f = 200 mm and sCMOS camera Dhyana BSI400 (Tucsen, Gaishan Town, China). In the flow system, a flow-through quartz UV cell (526UV0.25, FireflySci, Northport, NY, USA) with a 0.3 T rare-earth magnet and a conical-shaped magnetic field concentrator was used. An AL-1000 syringe pump (World Precision Instruments, Sarasota, FL, USA) was used to provide the flow in the flow cell at the required rate.