Fluorescence images were recorded on a DeltaVision Widefield microscope (Applied Precision) using a ×60 1.42 NA oil immersion objective, a Cy5 filter set (excitation: 650/13, emission: 684/24) for Alexa 647 fluorescence and a Cy7 filter set (excitation: 740/13, emission: 809/81) for NIR autofluorescence. The system is equipped with a Photometrics HQ camera. Images were recorded with 4 × 4 pixel binning and critical illumination to be able to detect the NIR autofluorescence signal. Bright field images were recorded along with fluorescence images. Image processing was performed using the open-source image analysis software Fiji.
Deltavision wide field microscope
Manufactured by Cytiva
The Deltavision Wide Field microscope is a high-performance imaging system designed for advanced cell and tissue analysis. It provides a wide field of view and high-resolution imaging capabilities, enabling researchers to capture detailed images of cellular structures and dynamics. The Deltavision Wide Field microscope is a versatile tool suitable for a range of life science applications.
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Lab products found in correlation
5 protocols using deltavision wide field microscope
1
Multimodal Fluorescence Microscopy Imaging
2
Visualizing Transfected HeLa Cells
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HeLa cells transfected with pEGFP-N3 based constructs (see S1 Table ) and plated in 8-well chamber slides (Ihbidi) were stained with MitoTracker® Deep Red and Hoechst 3342 prior to visualisation using a Deltavision Wide Field microscope (Applied Precision). The stage was encapsulated by an environmental chamber set to 37°C and 5% CO2 was supplied to the cells during visualisation.
3
Microinjection and Imaging Technique
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Preparation for microinjection was carried out with a Femtotips II microinjection needle (Eppendorf) and a gas pressure injection system were used to inject oil into the Stage 14 egg chambers [31 (link)]. Imaging was performed simultaneously with injection on a DeltaVision wide-field microscope (Applied Precision) using a 20 × 0.75 NA numerical aperture.
4
Immunofluorescence Staining of Cells
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Cells plated on coverslips/chamber slides/wells in preparation for immunofluorescence were first washed x3 with PBS. They were then fixed with 3% paraformaldehyde (PFA) in PBS pH 7.2 for 15 min prior to permeabilisation with 0.5% Triton-X, PBS pH 7.2 for 10 min and subsequent blocking with 2% BSA, 0.5% Triton-X, PBS pH 7.2 for 30 min. Incubations with primary antibody were performed in blocking buffer, typically at 1:100 dilution, overnight at 4°C. Alexa488 conjugated secondary antibodies were used at 1:750, again in blocking buffer for 1 h at 37°C. Coverslips were mounted onto glass slides using VectorShield medium with DAPI. MitoTracker® Deep Red staining of HeLa and primary fibroblasts was performed in complete media using 200 nM of probe for 20 min. Lysosomal staining was performed with 200 nM LysoTracker® Red for 15 min in complete media. Cells were subsequently washed x3 with PBS after which they were incubated in complete media for 30 min prior to fixation and visualisation. Images were acquired using a Deltavision Wide Field microscope and deconvoluted using Softworx software (Applied Precision).
5
Microinjection of Oil into Egg Chambers
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Preparation for microinjection was carried out with a Femtotips II microinjection needle (Eppendorf) and a gas pressure injection system were used to inject oil into the Stage 14 egg chambers [31] (link). Imaging was performed simultaneously with injection on a DeltaVision wide-field microscope (Applied Precision) using a 20x 0.75NA numerical aperture.
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