Nanoscope iiia controller

The hydrogel microstructure was visualized with atomic form microscopy (AFM) using a Veeco Multimode AFM (Veeco Instruments, Inc., Santa Barbara, CA, USA) with a Nanoscope IIIa controller. A 10 μL drop of the pentapeptide hydrogel (2% w/v) was transferred by pipetting onto a mica surface. After 30 s, the mica was rinsed with 300 μL Milli-Q water and air-dried. The height images were acquired using cantilevers with a spring constant of 10 N/m while the scan rate was adjusted to 1 Hz.
For transmission electron microscopy (TEM) visualization of the peptide hydrogel (2% w/v), an aliquot of 10 μL was deposited on carbon film-coated grids and air-dried prior imaging with TEM Jeol 2100 operated at 200 kV. ImageJ software was used to determine the mean fiber diameter by randomly measuring 100 fibers from the TEM images.

Scanning electron micrographs (SEM) were taken using a JEOL JSM-5510LV scanning electron microscope. The samples were mounted with adhesive conducting tape over an aluminium holder and sputtered with gold for 3 minutes (Agar Sputter Coater). The vacuum within the sputter unit was ca. 1 × 10⁻³ torr.
AFM images were recorded using a MultiMode atomic force microscope (NanoScope IIIa controller; Veeco). The stage was equipped with a video microscope to position the sample on the J scanner base. The samples were fixed to glass coverslips using sticky tabs over stainless steel sample holders. Before AFM analysis, the samples were observed on the metallic discs using a binocular GX reflective optical microscope equipped with a Motticam 2000 microscope digital camera. All images were recorded in tapping mode using TESP 15 series (HQ:NSC15/Al BS) sharpened silicon probes with nominal spring constant of 40 N m⁻¹ and nominal resonance frequency of 325 kHz (μmasch). The scan rate was changed according to the size of the scan area and the features observed on the surface. The scans were analysed using NanoScope software version 6.13 (Veeco). Each height image was processed using the plane-fitting third-order and the flatten zero-order commands in the software. For the amplitude images, the plane-fitting zero-order command was performed.

Atomic force micrographs (AFM) were acquired using a Digital Instruments Dimension™ 3000 SPM with a NanoScope IIIa controller (Veeco instruments, Santa Barbara, California, USA) and a NanoScope IIIa software (version 4.42r4). Tapping mode was employed across the surface of the samples at a frequency of 1 Hz.
The infrared (IR) spectra were recorded in the mid-infrared region (4000–550 cm⁻¹) on a Bruker ALPHA in diamond attenuated total reflection (ATR) using Opus software. The resolution was 2 cm⁻¹ at 128 scans.
Raman spectroscopy was performed on an iHR 320 model fully automated spectrometer with a wavelength position resolution of 0.2 cm⁻¹. The excitation source was an argon ion laser (514 nm) within the wavenumber range of 0–4000 cm⁻¹.

The entire volume of each sample was placed on freshly cleaved mica (1 × 1 cm). After 5 min, the solvent was absorbed with filter paper. MilliQ water (20 µL) was then placed on the mica surface and immediately absorbed with filter paper. This process was repeated three times to remove salts from the sample. All samples were dried in vacuo prior to AFM measurements. Tapping-mode images were obtained on a multimode scanning probe microscope with a Nanoscope IIIa controller (Veeco, Woodbury, NY, USA).

Atomic Force Microscopy was done using a Nanoscope IIIa controller (Veeco Instruments, Santa Barbara, CA, USA), silicon-nitride tip (NP-20, Veeco, k = 0.32 N/m) in the contact mode. The size of the scanned area was 10 × 10 μm (512 × 512 pixels). The AFM image analysis was done using Gwyddion 2.60 software (P. Klapetek, Czech Metrology Institute, Brno, Czech Republic and D. Nečas, University of Technology, Brno, Czech Republic) [18 (link)]. Before quantitative analysis, AFM images were corrected for polynomial background (second-order) to avoid the possible influence of the surface curvature. Also, the data were levelled by subtraction of the mean plane.

The samples BSA:chaperones A, B, C, D, E, F, and G (1:1) and BSA from kinetic assays at 65°C were diluted with milli-Q water in a 1:9 (v/v). 100 μL of the diluted sample were deposited on a mica and dried under vacuum over moisture-free silica gel. After drying micas, 500 μL of milli-Q water was added and allowed to stand for 5 min before removing the excess of water using filter paper. The micas were dried again for 24 hours and deposited over glass slides used in light microscopy. The samples were observed with an atomic force microscope (Bioscope, Digital Instruments, Santa Barbara CA, USA, equipped with a Nanoscope IIIa controller) working in contact mode with a silicon nitride probe (DNP, Veeco, USA). Microscope works on an inverted Diaphot 200 (Nikon) microscope with a 100 microns scanner and silicon nitride tips with 50 nm radius of curvature (DNP–Veeco). Sweep speeds used between 1969–1285 Hz, 10 nN force and 0.5 arbitrary units of gain.