Awareness for artifacts in fluorescence microscopy of β-TCP

Area of Science:

• Biomaterials Science
• Cell Biology
• Microscopy Techniques

Background:

• Fluorescence analysis of beta-tricalcium phosphate (β-TCP) ceramics is a common method for characterizing cell presence.
• Previously undescribed artifacts resembling cells have been observed, potentially leading to misinterpretation in fluorescence imaging.

Purpose of the Study:

• To identify and characterize novel artifacts observed during fluorescence analysis of β-TCP ceramics.
• To evaluate methods for reducing these artifacts to prevent false positive cell identification.

Main Methods:

• Investigated artifacts in β-TCP ceramics using fluorescence microscopy.
• Applied prolonged ultrasound washing and Technovit 9100 fixation as artifact reduction techniques.
• Assessed the efficacy of mechanical cleaning in conjunction with Technovit fixation.

Main Results:

• Undescribed artifacts with cell-like shapes and fluorescence properties were identified.
• Untreated β-TCP ceramics showed no reduction in artifacts regardless of further treatment.
• Technovit 9100 fixation significantly reduced artifacts, with further reduction achieved through mechanical cleaning.

Conclusions:

• Artifacts in β-TCP ceramics can be mistaken for cells, necessitating careful evaluation.
• Technovit fixation, especially when combined with mechanical cleaning, is an effective method for reducing these artifacts.
• Researchers should be aware of these artifacts and employ artifact reduction strategies to ensure accurate cell detection in β-TCP ceramic studies.