For transient expression of GFP, three combinations of the corresponding vectors were transfected into Pv11 cells as follows: 10 μg pPv121-MCS; 1 μg pPv121-Tet-On 3G and 9 μg pTetO-202bp-AcGFP1; 1 μg pPv121-MCS and 9 μg pPv121-AcGFP1. One day after transfection, the medium was replaced with IPL-41 or trehalose mixture with or without Dox at 1 μg/mL. After 24 h, the AcGFP1 fluorescence in the cells was viewed using a Biozero BZ-X700 microscope (Keyence, Osaka, Japan). AcGFP1 expression was evaluated by Western blot analysis.
Biozero bz x700 microscope
Manufactured by Keyence
Sourced in Japan
The Biozero BZ-X700 is a digital microscope designed for laboratory use. It features a high-resolution camera and advanced imaging capabilities to capture and analyze samples. The BZ-X700 provides clear, detailed images and can be used for a variety of research and analysis tasks.
Automatically generated - may contain errors
Lab products found in correlation
Cellstain double staining kit, by Dojindo Laboratories (1 mentions)
Pecam 1, by BD (1 mentions)
Prox1, by R&D Systems (1 mentions)
Cy5 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)
Lyve 1, by Abcam (1 mentions)
Tcs sp5 confocal microscope, by Leica (1 mentions)
Blocking reagent, by PerkinElmer (1 mentions)
4 protocols using biozero bz x700 microscope
1
Transient GFP Expression in Pv11 Cells
2
Pv11 Cell Viability and Proliferation
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After rehydration in fresh complete IPL-41 medium, Pv11 cells were collected at 1, 3, 5, and 7 d following rehydration and stained with a Cellstain Double Staining Kit (DOJINDO), following the manufacturer’s instructions. Bright-field images and calcein-AM fluorescence were visualized under a Biozero BZ-X700 microscope (Keyence), and Pv11 cells were quantified using BZ-H3C call count software (Keyence). One day after rehydration, cell viability was estimated from the number of calcein-AM–positive (live) cells, divided by the total number of cells observed by bright-field microscopy. Cell proliferation was assessed from the total number of cells counted every 2 d by bright-field microscopy. Each measurement was made with 12 replicates.
3
Transient GFP expression in Pv11 cells
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For transient expression of GFP, three combinations of the corresponding vectors were transfected into Pv11 cells as follows: 10 μg pPv121-MCS; 1 μg pPv121-Tet-On 3G and 9 μg pTetO-202bp-AcGFP1; 1 μg pPv121-MCS and 9 μg pPv121-AcGFP1 (Table S2 ). One day after transfection, the medium was replaced with IPL-41 or trehalose mixture with or without Dox at 1 μg/mL. After 24 h, the AcGFP1 fluorescence in the cells was viewed using a Biozero BZ-X700 microscope (Keyence, Osaka, Japan). AcGFP1 expression was evaluated by western blot analysis.
4
Immunofluorescence Analysis of Mouse Skin
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After mouse embryos were fixed with 4% paraformaldehyde (PFA)/PBS at 4 °C and subsequently dehydrated in methanol, the dorsal skin were dissected, rehydrated in PBST (0.2% Tween-20/PBS), and incubated in the blocking buffer consisting of 0.1 M Tris-HCl, pH7.5, 0.5% blocking reagent (PerkinElmer Life Sciences) and 0.15 M NaCl for 1 hour at room temperature (r.t.). The skin samples were incubated with the primary antibodies against LYVE-1 (Abcam), Prox1 (R&D Systems), PECAM-1 (BD Biosciences), or Ki67 (Abcam) at 4 °C overnight and subsequently with Alexa Fluor®-488-, Alexa Fluor®-546-, Alexa Fluor®-594-, Cy3-, and Cy5-conjugated secondary antibodies (Thermo Fisher Scientific). Immunofluorescence images were obtained with Biozero BZ-X700 microscope (Keyence, Japan) or Leica TCS SP5 confocal microscope. The branch number, total vessel length, and width of lymphatic vessels were analyzed by NIH ImageJ software.
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