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The BALTEC is a high-performance tabletop freeze dryer designed for laboratory use. It is a compact and versatile unit that can be used for the lyophilization of a wide range of samples, including biological materials, pharmaceuticals, and food products. The BALTEC features a powerful refrigeration system, programmable control, and efficient condensation capabilities to ensure consistent and reliable freeze drying results.

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6 protocols using baltec

1

Scanning Electron Microscopy of Cultured HCEnCs

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HCEnCs were seeded on FNC-coated glass coverslips and maintained in M5-Endo for at least 48 hours. Following a four-day storage, coverslips were removed and fixed with neutrally buffered 2% glutaraldehyde (Electron Microscopy Sciences, Hatfield, PA, USA) at 4 °C for 4 hours. Alternatively, samples were placed at 37 °C for 48 hours recovery, and subsequently fixed. Samples were processed as previously described17 (link). Briefly, samples were post-fixed in 1% osmium tetroxide at room temperature for 1 hour. The samples were then dehydrated, critical point dried (BALTEC, Balzer, Liechtenstein) and mounted onto a metal stub using carbon adhesive tabs. Samples were sputter-coated for 160 seconds with a 22 nm layer of gold-phalladium alloy (BALTEC) and examined under a scanning electron microscope (Quanta 650FEG; FEI, Hillsboro, OR, USA).
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2

Corneal Endothelial Denuded Area Analysis

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The endothelial denuded area along the laser trephination was evaluated with scanning electron microscopy (SEM). Corneas were fixed in neutral buffered 2% glutaraldehyde (Electron Microscopy Sciences, Hatfield, USA) at 4 °C for 24 h. After rinse with 1 × PBS, they were cut into halve and post-fixed in aqueous solution of 1% osmium tetroxide at room temperature for 30 min. The samples were then dehydrated under an increasing alcohol gradient: 25% ethanol for 5 min, 50% ethanol for 5 min, 75% ethanol for 5 min, 95% ethanol for 5 min, 3 × 100% ethanol for 10 min each, followed by critical point drying (BALTEC, Balzer, Liechtenstein). Dehydrated samples were then mounted onto a metal stub using carbon adhesive tabs. Samples were sputter-coated with a 25-nm layer of gold–palladium alloy (BALTEC), and examined under a scanning electron microscope (Quanta 650FEG; FEI, Hillsboro, OR). The denuded endothelial area, alongside the laser path for each cornea sample, was measured using ImageJ (National Institutes of Health, Bethesda, MD). The denuded area was defined as regions absent of endothelial cells from the laser cut.
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3

SEM Analysis of 3DP Scaffolds

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Using a Philips-XL-30 instrument (FEI, Hillsboro, OR, USA), the surface morphology of the 3DP scaffolds and 3DP with ESP collagen film scaffolds was analyzed by SEM. The samples were fixed in 2% glutaraldehyde for 2 h at 4 °C, washed twice in phosphate buffer solution (PBS), and post-fixed in 1% osmic acid for 2 h at 4 °C. After two PBS washes, the samples were dehydrated in a graded series of ethanol, dried to a critical point (HCP-2; Hitachi, Tokyo, Japan), mounted, sputter-coated with gold (BAL-TEC; FEI), and analyzed by SEM.
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4

Corneal Specimen Ultrastructure Analysis

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The corneal specimens obtained were examined by electron microscopy. First, the corneal specimens were immersed in a stationary liquid of 0.1 M sodium bicarbonate (pH 7.4; Electron Microscopy Science, Hatfield, PA, USA) containing 2.5% glutaraldehyde overnight at 4°C. Then, the corneal specimens were immersed in 1% osmium tetroxide (FMB, Singapore) at 22°C for 2 hours. Next, the corneal specimens were dehydrated in a series of ethanol diluents, dried in a critical point dryer (BALTEC, Balzers, Liechtenstein), and installed on a holder. At last, a 10 nm thick gold layer of (BALTEC) was sputtered on the specimens that were examined by scanning electron microscope (SEM; FEI, Holland).
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5

Scanning Electron Microscopy of Leishmania Cells

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Leishmania cells were layered on poly-lysin-coated coverslips, deshydrated in graded series of ethanol followed by critical point drying (CDP-030, BAL-TEC). Coverslips were mounted on specimen stubs, coated with 9 nm of platinum (SCD 050 sputter/coater BAL-TEC) and observed with FEI Quanta FEG200 microscope. All chemicals were from Electron Microscopy Sciences (Hatfield, PA, USA), solvents were from Sigma-Aldrich (Saint-Louis, MO, USA). Images were treated with Fiji and the figures were mounted using the FigureJ plugin.
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6

Scanning Electron Microscopy of Excised TE-EK Grafts

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Excised TE-EK graft specimens for scanning electron microscopy were first immersed overnight in a fixative solution consisting of 2% glutaraldehyde in PBS (pH 7.4; Electron Microscopy Sciences, Hatfield, PA, USA) at 4 °C overnight. Fixed specimen was washed thrice in PBS for 5 minutes each, and kept in 1% osmium tetroxide at room temperature for 1 hour. The sample was then dehydrated in an increasing concentration of ethanol at 25%, 50%, 75%, 95% and 100% for 10 minutes each, with the last step repeated thrice. Dehydrated samples were then dried in a critical point dryer (BALTEC, Balzer, Liechtenstein), and mounted onto a SEM stub using carbon adhesive tabs. Samples were then sputter-coated with a 10 nm layer of gold (BALTEC), and examined under a scanning electron microscope (Quanta 650FEG; FEI, Hillsboro, OR, USA).
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