Plan apochromat 20x 0.8 objective lens

The expression of the TIGAR protein and E6 viral oncoprotein within the HPV16-infected cervical cancer clinical samples by immunofluorescence staining, as well as the detection of cellular apoptosis using Annexin V-FITC/PI (or DAPI), were visualized confocal fluorescence-microscopy on a Zeiss LSM800 instrument using a Plan-Apochromat 20x/0.8 objective lens and ZEN OS software (Carl Zeiss Microscopy). The relative fluorescence-intensities of the Anti-TIGAR-specific (Rhodamine Red-X-positive) and Anti-HPV16 E6-specific (Alexa Fluor 488-positive) signals were graphically quantified using the Zen 2.5D analysis tool (Carl Zeiss Microscopy).

Carmine dyes such as carminic acid are used to complex with glycogen and short glucose polymers to produce a fluorescent signal responsive to the levels of glucose polymer. Thus, it is a useful tool to analyze glycogen storage [22 (link)]. Carminic acid was introduced to worms through feeding as previously described [22 (link)]. Briefly, OP50 was inoculated into LB broth with 1 mg/ml carminic acid and grown overnight at room temperature in a light protected vial. OP50 with carminic acid was then used to seed standard NGM plates. Animals were plated on carminic acid containing plates following initial mid-L4 harvest (controls) or post-ARD at day 5, day 10, and day 30 and were visualized with Zeiss LSM 700 confocal microscope using a Plan-Apochromat 20x/0.8 objective lens, as detailed below.