To assess clonogenic properties the samples of the unstained CD34+ cells, PKH26 and PKH67 stained CD34+ cells and fused (HHCC) were collected and analyzed at 3 hours and 7 days after fusion. Briefly, cells were cultured in separate flasks, 1×103 cells were plated in a 35 mm dishes following the MethoCult® (StemCell Technologies) manufacturer’s instructions in optimized methylcellulose-based medium (MethoCult® H4034 StemCell Technologies) supporting the optimal growth of the erythroid progenitors [CFU-E and burst forming unit (BFU)-E], granulocyte macrophage progenitors (CFU-GM, CFU-G and CFU-M) and the multi-potential granulocyte, erythroid, macrophage, megakaryocyte progenitors (CFU-GEMM). The cells were incubated for 3 hours and 7 days and then the number of colonies was scored under the light microscope (Leica DM 5500B Compound Microscope with Leica DFC290 Color Digital Camera, Leica, Germany).
Leica dfc290 color digital camera
Manufactured by Leica camera
Sourced in Germany
The Leica DFC290 is a color digital camera designed for microscopy applications. It features a high-resolution sensor and captures images with accurate color representation. The camera is capable of providing clear and detailed images of microscopic samples.
Automatically generated - may contain errors
Lab products found in correlation
Methocult h4034, by STEMCELL (1 mentions)
Dm5500b compound microscope, by Leica camera (1 mentions)
Methocult, by STEMCELL (1 mentions)
Spinning disk confocal microscope, by Quorum Technologies (1 mentions)
C9100 13 em ccd camera, by Hamamatsu Photonics (1 mentions)
Ix81 microscope, by Hamamatsu Photonics (1 mentions)
Leica dmi6000 microscope, by Leica camera (1 mentions)
M165 fc stereomicroscope, by Leica camera (1 mentions)
3 protocols using leica dfc290 color digital camera
1
Clonogenic Potential of CD34+ Cells
2
Lignin Staining of Plant Internodes
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For lignin staining, internode samples were collected at reproduction developmental stage (R1). Internode were cut by Vibratome series 3000 to generate 60 μm cross sections and cleared by ethanol overnight. Cleared sections were immersed in 1% chloroglucinol staining solution (in 2 : 1 ethanol/HCl) for 2 min (Baum, 2008 ; Bart et al., 2010 ; Wuddineh et al., 2015 ). The cross sections were placed on microscopy slides and covered by coverslip. The edges of the slides were sealed with commercial sealant and examined under Leica DM 5500B Compound Microscope with Leica DFC290 Color Digital Camera at 10×.
3
Quantitative Analysis of Endodermal Cell Dynamics
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WashU: For Figure 4 D-G, fluorescence images were collected on a spinning disk confocal microscope (Quorum, Canada) using an inverted Olympus IX-81 microscope, a Hamamatsu EMCCD camera (C9100-13) and Metamorph acquisition software. For Figures 2A,5, and 6, WISH images were collected on a Nikon Macroscope with a Nikon AZ100 objective, a 1x lens N.A. 0.1, and a 4x lens N.A. 0.4. Iowa: For still epifluorescence images, live or fixed embryos were mounted in 2% methylcellulose and photographed using a Leica DMI 6000 microscope with a 5´/NA 0.15 objective or a 10´/NA 0.3 objective. For ISH images in Figure 3, embryos were mounted in 80% glycerol/PBS and photographed using a Leica M165FC Stereomicroscope with a Leica DFC290 Color Digital Camera.
For time-lapse imaging of endodermal cells in Figure 2 C-G and Figure 4 A-C, Tg(sox17:EGFP) embryos were embedded in 0.8% low-melting agarose in a dorsal-mount imaging mold as previously described (Ye et al., 2015) . Time-lapse imaging was taken in the dorsal region of endoderm at 25°C, at 5-minute interval with a 5´/NA 0.15 objective on an inverted Leica DMI 6000 microscope. Images were processed and cell tracking analyzed using ImageJ (Schneider et al., 2012) . Data were exported to Excel where cell migration speed, paths, direction were determined as previously reported (Lin et al., 2005) .
For time-lapse imaging of endodermal cells in Figure 2 C-G and Figure 4 A-C, Tg(sox17:EGFP) embryos were embedded in 0.8% low-melting agarose in a dorsal-mount imaging mold as previously described (Ye et al., 2015) . Time-lapse imaging was taken in the dorsal region of endoderm at 25°C, at 5-minute interval with a 5´/NA 0.15 objective on an inverted Leica DMI 6000 microscope. Images were processed and cell tracking analyzed using ImageJ (Schneider et al., 2012) . Data were exported to Excel where cell migration speed, paths, direction were determined as previously reported (Lin et al., 2005) .
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