Blc. viridis photosynthetic membranes were immobilized on the mica substrate in 40 μl absorption buffer (10 mM Tris-HCl pH 7.2, 150 mM KCl, 25 mM MgCl2) for 1 h at room temperature. The sample was rinsed with imaging buffer (10 mM Tris-HCl pH 7.2, 150 mM KCl). AFM imaging was performed using a Bruker Multimode 8.0 equipped with a 97 μm J-scanner and OTR4-10 probe (spring constant = 0.08 N m−1) in PeakForce Quantitative Nanoscale Mechanical (PeakForce QNM) mode. Minimal loading forces of ~120 pN were used at scan frequencies of 3 Hz using optimized feedback parameters. High-speed AFM was carried out using a NanoWizard 3 AFM (JPK) equipped with an ULTRA S scanner and Ultra-Short Cantilever probe (0.3 N·m−1, Nano World) in AC mode with the scan frequency of 20–30 Hz. Image analysis was carried out using Gwyddion and ImageJ. Statistical data are presented as mean ± standard error of the mean (SEM) unless stated otherwise.
Ultra short cantilever probe
Manufactured by NanoWorld
The Ultra-Short Cantilever probe is a specialized scanning probe microscopy (SPM) tool designed for high-resolution imaging and analysis. Its core function is to provide a sensitive and precise measurement of surface topography and properties at the nanoscale level. The ultra-short cantilever design offers enhanced stability and responsiveness, enabling accurate and reliable data acquisition.
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2 protocols using ultra short cantilever probe
1
Atomic Force Microscopy of Photosynthetic Membranes
2
Atomic Force Microscopy of Thylakoid Membranes
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Two microliters of thylakoid membrane samples were adsorbed onto freshly cleaved mica surface with 38 μL of adsorption buffer (10 mM Tris-HCl, pH 7.5, 150 mM KCl, 25 mM MgCl 2 ) at room temperature for 1.5 h. After adsorption, the sample was carefully rinsed with 800 μL imaging buffer (10 mM Tris-HCl, pH 7.5, 150 mM KCl) 71 (link) . High-resolution imaging was performed in AC imaging mode in liquid at room temperature using a NanoWizard 3 AFM (JPK) equipped with an ULTRA S scanner and Ultra-Short Cantilever probe (0.3 N•m -1 , Nano World) at scan frequencies of 5 Hz using optimised feedback parameters and a resolution of 512 × 512 pixels. The tip spring constant was routinely calibrated. Images were processed with JPK SPM Data Processing (JPK) and ImageJ.
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