Graduate Category: Engineering and Technology Degree Level: PhD Abstract ID#: 80

Methods Discussion Abstract

Background

Conclusion

David Walsh, Juliette Kassas, Shashi K. Murthy

• ~2.3 million Americans suffer from uveitis, and vision-loss due to

uveitis accounts for 10-15% of the blindness in the U.S. [2].

• Primary intraocular lymphoma (PIOL) has a two-year survival rate

of 39% [3, 4].

• PIOL is commonly referred to as masquerade syndrome due to its

gradual onset and ability to mimic other ocular diseases [5].

• The current diagnostic approach analyzes a biopsy sample from the

vitreous humor.

• A definitive diagnosis is achieved by characterizing cells within the

eye; an invasion of T-lymphocytes implicates uveitis while B-

lymphocytes implicate PIOL.

• There is an urgent need to fill the technological gap for diagnosis of

idiopathic eye disease.

Research impacts:

• Patient can get diagnosis during visit

• Save money on materials, equipment, and lab tests

• Small footprint and portable (7.5 cm x 2.5 cm x 0.3 cm)

• No need for experienced lab technicians

• Preserves sample for further analysis

• Prevents scarring of eye tissue and permanent vision loss

• Determines accurate course of treatment

T- and B-lymphocytes within the vitreous biopsy are too similar to

be separated solely on physical properties. Chemotaxis provides a

label-free separation alternative. We have started the development of

a paper-based, chemotactic, cell immunophenotyping device. This

process bypasses cell transit from the clinic to the lab and additional

preprocessing steps, which can frequently impact cell integrity and

viability, compromising analysis. Implementation at the point-of-

care will rapidly diagnose uveitis and PIOL, improving overall

patient prognoses.

Improper diagnosis of idiopathic eye disease can have disastrous

results. Uveitis and primary intraocular lymphoma (PIOL) are

symptomatically similar diseases but require drastically different

treatments. Current methods have a diagnostic yield of only 20% [1].

We are developing a point-of-care platform to rapidly characterize cells

within the eye for definitive diagnosis. This method will minimize cost

and time, while improving fidelity of results. The implementation of

the system at the point-of-care will ensure proper treatment.

Current diagnostic approaches involve analysis by flow cytometry and histology.

PIOL is also known as

masquerade syndrome.

Current approach is limited in efficacy due to: 1) Low number of cells and 2) Fragility of sample

Solution: A label-free separation method that can be performed at the point-of-care (POC)

4. Document the migration path of each cell over a 20 minute period,

using CellProfiler software.

5. Calculate the chemotactic index (C.I.) as displacement towards

gradient (x) over total migration distance (d).

6. Determine if C.I. value indicates if cellular movement is due to

implementation of the gradient.

Device prototype

References: 1. Margolis, R., et al., Vitrectomy for the diagnosis and management of uveitis of unknown cause. Ophthalmology, 2007. 114(10): p. 1893-7.

2. Durrani, O.M., C.A. Meads, and P.I. Murray, Uveitis: a potentially blinding disease. Ophthalmologica, 2004. 218(4): p. 223-36.

3. Ferreri, A.J., et al., Relevance of intraocular involvement in the management of primary central nervous system lymphomas. Ann Oncol, 2002. 13(4): p. 531-8.

4. Schabet, M., Epidemiology of primary CNS lymphoma. J Neurooncol, 1999. 43(3): p. 199-201.

5. Korfel, A., et al., Das Masquerade-Syndrom. Dtsch Arztebl, 2007. 104(8): p. 490-5.

Acknowledgements: This material is based upon work supported by the National Science Graduate Research

Fellowship awarded to D.I.W. under grant number NSF/DGE-0946746. Any opinion, findings

and conclusions or recommendations expressed in this material of those of the author(s) and do

not necessarily reflect the views of the National Science Foundation

C. I.x

d

*Lin, F., et al., Lab Chip. 2006.

*CellProfiler was developed by the Broad Institute

BCA-1 SDF-1α

Attracts B lymphocytes

(PIOL)

Attracts T lymphocytes

(Uveitis)

1. Collect patient sample via pars plana vitrectomy.

2. Establish a chemotactic gradient and place cells on the testing

platform.