We propose and demonstrate a light-sheet-based 3D interrogation system on a microfluidic platform for screening biological specimens during flow. To achieve this, a diffraction-limited light-sheet (with a large field-of-view) is employed to optically section the specimens flowing through the microfluidic channel. This necessitates optimization of the parameters for the illumination sub-system (illumination intensity, light-sheet width, and thickness), microfluidic specimen platform (channel-width and flow-rate), and detection sub-system (camera exposure time and frame rate). Once optimized, these parameters facilitate cross-sectional imaging and 3D reconstruction of biological specimens. The proposed integrated light-sheet imaging and flow-based enquiry (iLIFE) imaging technique enables single-shot sectional imaging of a range of specimens of varying dimensions, ranging from a single cell (HeLa cell) to a multicellular organism (C. elegans). 3D reconstruction of the entire C. elegans is achieved in real-time and with an exposure time of few hundred micro-seconds. A maximum likelihood technique is developed and optimized for the iLIFE imaging system. We observed an intracellular resolution for mitochondria-labeled HeLa cells, which demonstrates the dynamic resolution of the iLIFE system. The proposed technique is a step towards achieving flow-based 3D imaging. We expect potential applications in diverse fields such as structural biology and biophysics.

1.
F. C.
Zanacchi
,
Z.
Lavagnino
,
M. P.
Donnorso
,
A. D.
Bue
,
L.
Furia
,
M.
Faretta
, and
A.
Diaspro
,
Nat. Methods
8
,
1047
(
2011
).
2.
F. C.
Zanacchi
,
Z.
Lavagnino
,
M.
Faretta
,
L.
Furia
, and
A.
Diaspro
,
PLoS One
8
,
e67667
(
2013
).
3.
K.
Mohan
and
P. P.
Mondal
,
Rev. Sci. Intrum.
87
,
066107
(
2016
).
4.
K.
Mohan
and
P. P.
Mondal
,
Appl. Phys. Lett.
106
,
083112
(
2015
).
5.
L.
Samsel
,
P. K.
Dagur
,
N.
Raghavachari
,
C.
Seamon
,
G. J.
Kato
, and
J. P.
McCoy
, Jr.
,
Cytometry Part B.
84
,
379
(
2013
).
6.
C.
De la Calle
,
P. E.
Joubert
,
H. K. W.
Law
,
M.
Hasan
, and
M. L.
Albert
,
Autophagy
7
,
1045
(
2011
).
7.
N. A.
Bakhtina
and
J. G.
Korvink
,
RSC Adv.
4
,
4691
(
2014
).
8.
S.
Mondal
,
E.
Hegarty
,
C.
Martin
,
S. K.
Gokce
,
N.
Ghorashian
, and
A. B.
Yakar
,
Nat. Commun.
7
,
13023
(
2016
).
9.
N. S.
Barteneva
,
E. F.
Kan
, and
I. A.
Vorobjev
,
J. Histochem. Cytochem.
60
,
723
(
2012
).
10.
Y.
Han
,
Y.
Gu
,
A. C.
Zhang
, and
Y. H.
Lo
,
Lab Chip
16
,
4639
(
2016
).
11.
J. R.
Heath
,
Proc. Natl. Acad. Sci.
112
,
14436
(
2015
).
12.
P.
Horvath
,
N.
Aulner
,
M.
Bickle
,
A. M.
Davies
,
E. D.
Nery
,
D.
Ebner
,
M. C.
Montoya
,
P.
stling
,
V.
Pietiinen
,
L. S.
Price
,
S. L.
Shorte
,
G.
Turcatti
,
C. V.
Schantz
, and
N. O.
Carragher
,
Nat. Rev. Drug Discovery
15
,
751
(
2016
).
13.
B.
Patra
,
C. C.
Peng
,
W. H.
Liao
,
C. H.
Lee
, and
Y. C.
Tung
,
Sci. Rep.
6
,
21061
(
2016
).
14.
R.
Regmi
,
K.
Mohan
, and
P. P.
Mondal
,
Microsc. Res. Tech.
76
,
1101
(
2013
).
15.
R.
Regmi
,
K.
Mohan
, and
P. P.
Mondal
,
AIP Adv.
4
,
097125
(
2014
).
16.
S.
Quint
,
A. F.
Christ
,
A.
Guckenberger
,
S.
Himbert
,
L.
Kaestner
,
S.
Gekle
, and
C.
Wagner
,
Appl. Phys. Lett.
111
,
103701
(
2017
).
17.
B. B.
Collier
,
S.
Awasthi
,
D. K.
Lieu
, and
J. W.
Chan
,
Sci. Rep.
5
,
10751
(
2015
).
18.
Z.
Darzynkiewicz
,
E.
Bedner
,
X.
Li
,
W.
Gorczyca
, and
M. R.
Melamed
,
Exp. Cell Res.
249
,
1
(
1999
).
19.
R.
Wollman
and
N.
Stuurman
,
J. Cell Sci.
120
,
3715
(
2007
).
20.
J.
Wu
,
J.
Li
, and
R. K. Y.
Chan
,
Opt. Exp.
21
,
14474
(
2013
).
21.
H.
Deschout
,
K.
Raemdonck
,
S.
Stremersch
,
P.
Maoddi
,
G.
Mernier
,
P.
Renaud
,
S.
Jiguet
,
A.
Hendrix
,
M.
Bracke
,
R. V. D.
Broecke
,
M.
Rding
,
M.
Rudemo
,
J.
Demeester
,
S. C. D.
Smedt
,
F.
Strubbe
,
K.
Neyts
, and
K.
Braeckmans
,
Nanoscale
6
,
1741
(
2014
).
22.
P.
Pai
,
F.
Bragheri
,
A.
Bassi
, and
R.
Osellame
,
Lab Chip
16
,
1556
(
2016
).
23.
M. B.
Bouchard
,
V.
Voleti
,
C. S.
Mendes
,
C.
Lacefield
,
W. B.
Grueber
,
R. S.
Mann
,
R. M.
Bruno
, and
E. M. C.
Hillman
,
Nat. Photonics
9
,
113
119
(
2015
).
24.
A. K. S.
Lau
,
H. C.
Shum
,
K. K. Y.
Wong
, and
K. K.
Tsia
,
Lab Chip
16
,
1743
(
2016
).
25.
E. J.
Gualda
,
H.
Pereira
,
T.
Vale
,
M. F.
Estrada
,
C.
Brito
, and
N.
Moreno
,
Biomed. Opt. Express
6
,
4447
(
2015
).
26.
D. A.
Basiji
,
W. E.
Ortyn
,
L.
Liang
,
V.
Venkatachalam
, and
P.
Morrissey
,
Clinics Lab. Med.
27
,
653
(
2007
).
27.
K.
Goda
,
K. K.
Tsia
, and
B.
Jalali
,
Nature
458
,
1145
(
2009
).
28.
K.
Goda
,
A.
Ayazi
,
D. R.
Gossett
,
J.
Sadasivam
,
C. K.
Lonappan
,
E.
Sollier
,
A. M.
Fard
,
S. C.
Hur
,
J.
Adam
,
C.
Murray
,
C.
Wang
,
N.
Brackbilla
,
D. D.
Carlo
, and
B.
Jalali
,
Proc. Natl. Acad. Sci.
109
,
11630
(
2012
).
29.
E. D.
Diebold
,
B. W.
Buckley
,
D. R.
Gossett
, and
B.
Jalali
,
Nat. Photonics
7
,
806
(
2013
).
30.
A. H.
Voie
,
D. H.
Burns
, and
F. A.
Spelman
,
J. Microsc.
170
,
229
(
1993
).
31.
C. J.
Engelbrecht
and
E. H. K.
Stelzer
,
Opt. Lett.
31
,
1477
(
2006
).
32.
P. J.
Keller
,
A. D.
Schmidt
,
J.
Wittbrodt
, and
E. H. K.
Stelzer
,
Science
322
,
1065
(
2008
).
33.
E. H. K.
Stelzer
,
Nat. Methods
12
,
23
(
2015
).
34.
W. H.
Richardson
,
J. Opt. Soc. Am. A
62
,
55
(
1972
).
35.
L. B.
Lucy
,
Astron. J.
79
,
745
(
1974
).
36.
P. P.
Mondal
,
G.
Vicidomini
, and
A.
Diaspro
,
J. Appl. Phys.
102
,
044701
(
2007
).
37.
J.
Huisken
,
J.
Swoger
,
F.
Del Bene
,
J.
Wittbrodt
, and
E. H. K.
Stelzer
,
Science
305
,
1007
(
2004
).
38.
S. E.
Hulme
,
S. S.
Shevkoplyas
,
J.
Apfeld
,
W.
Fontana
, and
G. M.
Whitesides
,
Lab Chip
7
,
1515
(
2007
).
39.
C. K.
Rasmi
,
K.
Rajan
, and
P. P.
Mondal
,
Microsc. Res. Tech.
78
,
331
(
2015
).
40.
A.
Jabbar
,
C. K.
Rasmi
,
S.
Dilipkumar
,
K.
Rajan
, and
P. P.
Mondal
,
AIP Adv.
5
,
084801
(
2015
).
41.
Q.
Shan
,
J.
Jia
, and
A.
Agarwala
,
ACM Trans. Graphics
27
,
73
(
2008
).
42.
K.
Palikaras
,
E.
Lionaki
, and
N.
Tavernarakis
,
Nature
521
,
525
(
2015
).
43.
M. B.
Ahrens
,
M. B.
Orger
,
D. N.
Robson
,
J. M.
Li
, and
P. J.
Keller
,
Nat. Methods
10
,
413
(
2013
).

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