Sartorius eBook - 25

THE FUTURE OF DRUG DISCOVERY: LIVE 3D CELL ANALYSIS

into dendrites and axons over extended

in vitro models of neurodegenerative

timeframes, therefore, neuronal assays can take

conditions, such as Alzheimer's

weeks or even months-as in the case of stem

Disease and Huntington's Disease, in addition

cells-to develop and manipulate. By relying

to allowing the investigation of factors that

on predetermined endpoints with conventional

impact neuronal development6-8.

techniques, it is possible that crucial biological

Using label-free multiparametric

events could be missed or inaccurately

measurements, live-cell imaging and analysis can

determined. Live-cell imaging is a powerful

quantify neurite development (length, branch

monitoring tool for capturing these subtle

points), cell migration, and proliferation. Together

morphological fluctuations that occur during

these data can be used to characterize and

neuronal development and deterioration,

confirm differentiation stages over time (Figure

as well as the effects of active drugs. As an

2). Combining phase-contrast imaging with a

essential QC method, it provides unparalleled

non-perturbing nuclear-restricted fluorophore

spatial and temporal insight when developing

delivers an extra level of insight into the cell-cycle

Phase-contrast imaging and nuclear fluorescent labeling dynamically monitor neurite outgrowth and
cell division over time
A.

C.

D.

Nuclear count

Neurite length

(1/mm2)

(mm/mm2)

2500

Undifferentiated

2000

20

Differentiated

15

1500
Differentiated

1000

5

500
0

B.

10

0

96

Time (h)

192

288

0
192

Undifferentiated
240

Time (h)

288

336

Figure 2: Visualization of SH-SY5Y neuroblastoma differentiation using phase-contrast
imaging coupled with nuclear count (Nuclight-Orange). Stably transfected SH-SY5Y
neuroblastomas were differentiated with gradual serum starvation (10%-0%)
combined with atRA (10 μM) and BDNF (50 ng/mL) and monitored for 14 days using
the Incucyte®. Phase and fluorescent images of (A) undifferentiated or (B) differentiated
neurons. Note the greater confluence and lower neurite levels of undifferentiated cells.
(C) Analysis of cell counts shows that cell division is lower in differentiated cells. (D)
Analysis of neurite outgrowth shows that neurite length is greater in differentiated
cells.

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