The tissue construct samples were fixed in formalin and processed in different chemicals to prepare specimens for transmission electron microscope (TEM) imaging [21 (link)]. The samples were rinsed in PBS, fixed in 1% osmium tetroxide for 1 h, washed in distilled water, and then dehydrated through graded ethanol. For epoxy resin infiltration, the samples were treated with an ethanol-acetone mixture (1:1, by volume) for 10 min, acetone for 10 min, acetone-epoxy resin (2:1, by volume) for 30 min, acetone-epoxy resin (1:1, by volume) for 30 min, acetone-epoxy resin (1:2, by volume) for 30 min, and epoxy resin overnight. The samples were then embedded in fresh epoxy resin and cured overnight at 60 °C. The embedded samples were sectioned and collected on copper grid for imaging with a TEM instrument (Jeol, Japan).
Tem instrument
Manufactured by JEOL
Sourced in Japan
The TEM (Transmission Electron Microscope) instrument is a specialized laboratory equipment designed for the high-resolution imaging and analysis of small-scale structures and materials. It operates by transmitting a beam of electrons through a thin sample, creating an enlarged and detailed image that can be observed and studied.
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3 protocols using tem instrument
1
Tissue Samples Preparation for TEM Imaging
2
Characterization of Nanoparticle Synthesis
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The bath sonicator (100 W, 42 kHz) and magnetic stirrer and heater were purchased from Fisher Scientific Co. Ltd. (Shanghai, China). The TEM instrument came from JEOL Ltd. (Welwyn Garden City, UK). The EDAX analysis was obtained using a JEOL JSM 6390 LA Analytical device (Tokyo, Japan). The UV-Vis spectrophotometer was from Thermo Scientific GENESYS 10S (Toronto, Canada). The FT-IR spectra were collected in the attenuated total reflectance (ATR) mode using a PerkinElmer RX FT-IR 2× instrument with diamond ATR and DRIFT attachment from PerkinElmer (Buckinghamshire, UK). The thermogravimetric analyser was from TA Instruments (New Castle, DE, USA).
3
Hybrid Mycobacterial Exosome Preparation
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The TEX, DCMV, and MPLA were mixed at a ratio of 5: 5: 1 (TEX’ protein: DCMV’ protein: MPLA). After incubating in a 45°C water bath for 30 min, the mixture was then vortexed and sonicated for 5 min. Subsequently, the suspension was physically extruded 11 times through 0.8 µm, 0.4 µm, and 0.2 µm polycarbonate membrane, using a miniextruder (Avanti, USA), to obtain uniform Hy‐M‐Exo. Free MPLA was purified using Nanosep (MWCO = 300 K). To quantify the amount of MPLA within Hy‐M‐Exo, a LAL assay was performed, according to the manufacturer's protocol. The morphology of Hy‐M‐Exo was observed through TEM. First, Hy‐M‐Exo was diluted in PBS. Each sample was deposited onto an EM grid and observed under a TEM instrument (JEOL, Akishima, Japan) at 100 kV. Zetasize Nano ZS (Malvern Instruments, Malvern, UK) was used to test the size distribution and the zeta potential.
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