Significance

Traditional pathology workflow suffers from limitations including biopsy invasiveness, small fraction of large tissue samples being analyzed, and complex and time-consuming processing.

Aim

We address limitations of conventional pathology workflow through development of a laser microbiopsy device for minimally invasive harvest of sub-microliter tissue volumes. Laser microbiopsy combined with rapid diagnostic methods, such as virtual hematoxylin and eosin (H&E) imaging has potential to provide rapid minimally invasive tissue diagnosis.

Approach

Laser microbiopsies were harvested using an annular shaped Ho:YAG laser beam focused onto the tissue surface. As the annulus was ablated, the tissue section in the center of the annulus was ejected and collected directly onto a glass slide for analysis. Cryogen spray cooling was used before and after laser harvest to limit thermal damage. Microbiopsies were collected from porcine skin and kidney. Harvested microbiopsies were imaged with confocal microscopy and digitally false colored to provide virtual H&E images.

Results

Microbiopsies were successfully harvested from porcine skin and kidney. Computational and experimental results show the benefit of cryogen pre- and post-cooling to limit thermal damage. Virtual H&E images of microbiopsies retained observable cellular features including cell nuclei.

Conclusions

Laser microbiopsy with virtual H&E imaging shows promise as a potential rapid and minimally invasive tool for biopsy and diagnosis.

Introduction: This study aimed at answering three research questions: (1) Under the experimental conditions studied, what is the dominant mechanism of Holmium:YAG lithotripsy with or without pulse modulation? (2) Under what circumstances can laser pulse modulation increase crater volume of stone ablation per joule of emitted radiant energy? (3) Are BegoStone phantoms a suitable model for laser lithotripsy studies? Materials and Methods: The research questions were addressed by ablation experiments with BegoStone phantoms and native stones. Experiments were performed under three stone conditions: dry stones in air, hydrated stones in air, and hydrated stones in water. Single pulses with and without pulse modulation were applied. For each pulse mode, temporal profile, transmission through 1 mm water, and cavitation bubble collapse pressures were measured and compared. For each stone condition and pulse mode, stones were ablated with a fiber separation distance of 1 mm and crater volumes were measured using optical coherence tomography. Results: Pulses with and without pulse modulation had high (>80%) transmission through 1 mm of water. Pulses without pulse modulation generated much higher peak pressures than those with pulse modulation (62.3 vs 11.4 bar). Pulse modulation resulted in similar or larger craters than without pulse modulation. Trends in BegoStone crater volumes differed from trends in native stones. Conclusions: This results of this study suggest that the dominant mechanism is photothermal with possible photoacoustic contributions for some stone compositions. Pulse modulation can increase ablation volume per joule of emitted radiant energy, but the effect may be composition specific. BegoStones showed unique infrared ablation characteristics compared with native stones and are not a suitable model for laser lithotripsy studies.

[This corrects the article DOI: 10.1117/1.JBO.27.12.125001.].