Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

A simple and stable auto focusing protocol for long multidimensional live cell microscopy.

F Wolf1, S Geley

  • 1Biocenter, Division of Molecular Pathophysiology, Innsbruck Medical University, Innsbruck, Austria.

Journal of Microscopy
|January 28, 2006
PubMed
Summary

Automated microscopy focus routines can harm live cells. This study introduces a new method using fluorescent markers to maintain focus, minimizing light exposure and improving live cell imaging safety.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Ultrahypofractionation of localized prostate cancer-an updated statement of the prostate cancer working group of the German Society of Radiation Oncology (DEGRO).

Strahlentherapie und Onkologie : Organ der Deutschen Rontgengesellschaft ... [et al]·2026
Same author

Dosimetric robustness of urethra sparing in ultrahypofractionated prostate SBRT using catheter-based motion tracking.

Strahlentherapie und Onkologie : Organ der Deutschen Rontgengesellschaft ... [et al]·2026
Same author

Prediction of Mutations and Outcome in Gastrointestinal Stromal Tumors with Deep Learning: A Multicenter, Multinational Study.

medRxiv : the preprint server for health sciences·2026
Same author

Biomarkers in prostate cancer: current status and future directions in radiotherapy-statement from the Prostate Cancer Working Group of the German Society of Radiation Oncology (DEGRO).

Strahlentherapie und Onkologie : Organ der Deutschen Rontgengesellschaft ... [et al]·2025
Same author

The risk of second malignancies following prostate cancer radiotherapy in the era of conformal radiotherapy: a statement of the Prostate Cancer Working Group of the German Society of Radiation Oncology (DEGRO).

Strahlentherapie und Onkologie : Organ der Deutschen Rontgengesellschaft ... [et al]·2024
Same author

Prostate cancer and elective nodal radiation therapy for cN0 and pN0-a never ending story? : Recommendations from the prostate cancer expert panel of the German Society of Radiation Oncology (DEGRO).

Strahlentherapie und Onkologie : Organ der Deutschen Rontgengesellschaft ... [et al]·2024

Area of Science:

  • Microscopy and Imaging Science
  • Cell Biology
  • Biophotonics

Background:

  • Focus maintenance is critical but challenging in multidimensional wide-field microscopy.
  • Conventional autofocus routines increase light exposure and can cause phototoxicity in live cells.
  • Automated focusing may erroneously target high-contrast artifacts, compromising image quality.

Purpose of the Study:

  • To develop an improved autofocus method for multidimensional microscopy.
  • To minimize light exposure and phototoxicity during live cell imaging.
  • To prevent focusing on irrelevant high-contrast objects.

Main Methods:

  • A novel technique was developed by labeling a specific position with a fluorescent marker.
  • This marker allows separation of the focusing procedure from actual image acquisition points.

Related Experiment Videos

  • A Visual Basic-based program was created to run before each image acquisition in multidimensional experiments.

    • Main Results:

    • The new method effectively separates focusing from image acquisition, overcoming limitations of routine autofocus.
    • This approach allows precise focus control while significantly reducing light exposure.
    • The technique minimizes light toxicity, making it suitable for sensitive live cell imaging.

    Conclusions:

    • Labeling a defined position with a fluorescent marker offers a superior autofocus strategy.
    • This method enhances focus stability in multidimensional microscopy.
    • It significantly reduces phototoxicity, enabling safer and more effective live cell imaging experiments.