We used the DIC detection mode of inverted microscope (IX-73; Olympus, Tokyo, Japan) equipped with a microscopic objective (LUCPlanFL N, NA = 0.7, 60×) and a color-sensitive camera (DP74; Olympus) (46 (link)). Light sources (U-LH100L and U-HGLGPS; Olympus) were used for DIC and fluorescence detection, respectively. The excitation wavelength was 470–495 nm, and the fluorescence at 510–550 nm was detected. The sample solution was cast on a coverslip (Matsunami Glass) and covered with a glass slide (Matsunami Glass). The coverslip and slide glass were cleaned with ethanol and 5 M KOH and coated with 0.5% 2-methacryloyloxyethyl phosphorylcholine (MPC) polymer (Lipidureμ-CM5206; NOF Corp.) in ethanol before use (47 ). DIC and fluorescence images were obtained at 21–22°C. The uptake ratio of DNA-binding proteins inside to outside DNA droplets was calculated from the ratios of the fluorescence intensities inside to outside DNA droplets.
U lh100l
Manufactured by Olympus
The U-LH100L is an LED illuminator designed for Olympus microscopes. It provides bright, uniform illumination for observation and imaging tasks.
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Lab products found in correlation
2 protocols using u lh100l
1
Microscopic Analysis of DNA Droplet Protein Uptake
2
Reflection Mode Microscopy Setup
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Our setup for collecting the training and testing data for all experiments is composed as a simple reflection mode microscope system (refer to Fig. 1c for the general schematic). We use an Olympus IX81 optical microscope as the centerpiece for our setup. The optical microscope features multiple ports and attachments, including an input reflection mode optical chamber to attach a light source to, and two output chambers that are connected to an attachable camera and optical fiber, respectively. The software and hardware setup allow the optical output of the microscope to be controlled between three paths: the first to the eyepiece for direct visualization, the second to the attached output camera chamber, and the third to the output optical fiber connected to a benchtop spectrometer. In our setup, we used a Hamamatsu ORCA-03G grayscale camera and an Ocean Optics Jaz visual spectrometer at the mentioned output ports. For the incident light, we use a pE-4000 cool LED for selective narrowband illumination, and a halogen lamp (Olympus U-LH100L) coupled to a liquid crystal tunable filter for broadband illumination. Chips with the 1D and 2D grating structures were placed on the microscope stage, with their orientation facing downward for analyzing their reflection mode. A ×4 objective lens (NA = 0.13) was used to analyze the structures and their resonance patterns.
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