On-line flow cytometry for real-time surgical guidance.
OBJECTIVE
This study tests the feasibility of using on-line analysis of tissue during surgical resection of brain tumors to provide biologically relevant information in a clinically relevant time frame to augment surgical decision making. For the purposes of establishing feasibility, we used measurement of deoxyribonucleic acid (DNA) content as the end point for analysis.


METHODS
We investigated the feasibility of interfacing an ultrasonic aspiration (USA) system with a flow cytometer (FC) capable of analyzing DNA content (DNA-FC). The sampling system design, tissue preparation requirements, and time requirements for each step of the on-line analysis system were determined using fresh beef brain tissue samples. We also compared DNA-FC measurements in 28 nonneoplastic human brain samples with DNA-FC measurements in specimens of 11 glioma patients obtained from central tumor regions and surgical margins after macroscopically gross total tumor removal to estimate the potential for analysis of a biological marker to influence surgical decision making.


RESULTS
With minimal modification, modern FC systems are fully capable of real-time, intraoperative analysis of USA specimens. The total time required for on-line analysis of USA specimens varies between 36 and 63 seconds; this time includes delivery from the tip of the USA to complete analysis of the specimen. Approximately 60% of this time is required for equilibration of the DNA stain. When compared with values for nonneoplastic human brain samples, 50% of samples (10 of 20) from macroscopically normal glioma surgical margins contained DNA-FC abnormalities potentially indicating residual tumor.


CONCLUSIONS
With an interface of existing technologies, DNA content of brain tissue samples can be analyzed in a meaningful time frame that has the potential to provide real-time information for surgical guidance. The identification of DNA content abnormalities in macroscopically normal tumor resection margins by DNA-FC supports the practical potential for on-line analysis of a tumor marker to guide surgical resections. The development of such a device would provide neurosurgeons with an objective method for intraoperative analysis of a clinically relevant biological parameter that can be measured in real time.
Mesiwala
Ali H
AH
Department of Neurological Surgery, University of Washington, Seattle, Washington, USA.
Scampavia
Louis D
LD
Rabinovitch
Peter S
PS
Ruzicka
Jaromir
J
Rostomily
Robert C
RC
D000782:Aneuploidy; D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis / Q000235:genetics; D001707:Biopsy, Needle / Q000295:instrumentation; D001921:Brain / Q000473:pathology / Q000601:surgery; D001932:Brain Neoplasms / Q000235:genetics / Q000473:pathology / Q000601:surgery*; D002417:Cattle; D004273:DNA, Neoplasm / Q000032:analysis*; D004453:Echoencephalography / Q000295:instrumentation; D005240:Feasibility Studies; D005434:Flow Cytometry / Q000295:instrumentation*; D005910:Glioma / Q000235:genetics / Q000473:pathology / Q000601:surgery*; D006801:Humans; D008856:Microscopy, Fluorescence; D038361:Neuronavigation / Q000295:instrumentation*; D009862:Online Systems / Q000295:instrumentation*; D011379:Prognosis; D014584:User-Computer Interface
D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm