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0:00
Hello. Today we are talking
about flow cytometry in general,
but more specifically on
systematic differences
between polychromatic
flow cytometry
and spectral flow cytometry,
which has been a
recent development.
My name is Martin Buescher.
I'm a physicist by training,
but I got my PhD in
Applied biotechnology.
I'm running the
BioPhysics group within
the Miltenyi company
that manufacturers
flow cytometry devices
and many other tools and
systems around immunology
and science in general.
0:36
First, I will briefly
introduce flow cytometry from
a historical point of view.
After that's done, I'm
going to talk about
the fundamental principles
that make up flow cytometry.
From there, some practical
consequences are
derived that play a role in
the use of flow cytometry.
From there, we're
going to discuss some
practical consequences in
real-life experiments,
and from that, we'll
deduce best practices
in daily work with
those devices.
Then we are almost through and
doing some final conclusions.
1:11
Introduction to flow cytometry.
Flow cytometry was invented
in the last century, in the 60s,
mainly by Wolfgang Dittrich
and Wolfgang Gohde.
They published a
paper on impulse
fluorometry of single
cells in suspension,
where they used
the light source,
excited fluorescence, and worked
out the fundamental
principle of flow cytometry,
which I'm going to talk
about in a minute.
Then in the 70s,
this whole technology had
developed quite a bit
further and spear-headed
by Leonard Herzenberg.
They not only introduced
cell sorting,
and patented the
technology by that,
they also developed
the systems further
so that they could be
used in routine labs.
Then in the 80s, starting
then until today,
the systems have developed
further and further
and got more complex with
more channels and are faster.
Some bigger examples are
publications from the mid-80s,
where they did 8-color, 10-parameter
flow cytometry experiments.
By that, they could preserve
a very different
differentiated image
of leukocyte heterogeneity
in the blood.
After that, things have
developed further.
Around the millennium,
17-color flow
cytometry experiments
have been conducted.
More or less, the
whole immune system,
as it is known today,
has been unraveled.
Now the question is
how to move from there
and how to expand it further.
This is probably with
spectral flow cytometry.
