Human Cell Culture & Live-Cell Imaging Study

Project Overview

Under the supervision of Dr. Mitra Tabatabaee, this human cell-culture study focused on establishing reliable culture conditions for U118-MG glioblastoma cells from the ground up. The project demanded sequential milestones—early recovery, consistent passaging, achievement of robust confluence, and downstream work that depended on stable cellular health and timing.

Project Journey

Animation shows: healthy cells → fungal contamination → sanitization → recovery with fluorescent imaging

Challenge: Fungal Contamination

The first major challenge emerged during early passages, which repeatedly failed to reach confluence. Microscopic inspection ultimately clarified the situation: fungal contamination was present. The contamination event did not occur in isolation—concurrently, the incubator's CO₂ supply dropped below required levels, disrupting bicarbonate buffering and elevating media pH, which introduced additional cellular stress.

This convergence of events underscored that cell culture outcomes are not determined solely by direct actions at the bench. Infrastructure is a component of the experiment, and deviations in environmental control can amplify the impact of biological contamination.

Resolution: Deep Sanitization

To overcome these setbacks, I led a comprehensive decontamination of the cell-culture laboratory with close attention to both visible and subtle sources of contamination risk. In parallel, the incubator environment was stabilized by installing a new carbon dioxide tank and re-establishing conditions appropriate for sensitive human cultures.

Strict aseptic protocols were implemented going forward as a reinforced operating standard, including more deliberate attention to workflow discipline and consistent environmental monitoring. Within four weeks, a fresh batch of U118-MG cells achieved approximately 80 percent confluence.

Live-Cell Imaging

By 96 hours post-transfection, approximately 15–20 percent of cells exhibited faint but discernible fluorescence, enabling visualization of actin cytoskeletal remodeling in live U118-MG cells. Although time constraints limited completion of glutamate chemotaxis assays, these preliminary imaging data established feasibility for monitoring live-cell morphological dynamics within the system.

Key Learnings

• •Microscopic inspection is an essential diagnostic tool, not merely a routine step
• •Aseptic technique and incubator stability must be treated as equally foundational
• •Decontamination resolves sterility, but cannot restore lost time—timelines must account for recovery
• •Resilience and systematic troubleshooting are applied skills that determine project momentum