So, that decade really starts in the late 60s to 1980, was a
marvelous time for cell biology. I mean, this work came out of
that time was a revolution for understanding how synaptic
transmission works. There was just an era of great cell
biology. So why do you think that happened?
And why did it end?
Well I don't think it did end.
Are you kidding me? It has just started up again.
The focus, you know, is molecular biology sort of swept away.
Oh yeah, I mean there was a period of time in the summer, of course,
you get these guys at Caltech. In fact, we had a student and
she got up and she said, well I want everybody to know that
electron microscopy is a dying field. And so we wanted
-- we made a plot of what a dying field looked like over our door.
A dying electromagnetic field. Remember that?
No!
It just showed this thing where all of a sudden the curve went
down. And it said "Dying Field." She was in Keith Porter's lab, too.
Really?
The founder of modern electron microscopy had become disillusioned.
And so there was a period of time when people thought that
we were going to understand everything by understanding genes.
And we've understood a lot from genes, but you don't understand everything
from genes. Now, finally, the technical side of electron microscopes
is having a drastic -- and new kinds of electron microscopes
are appearing. And the challenge is on people like John and me,
people who actually prepare stuff for electron microscopes
to take advantage of these new kinds of optical things that these
microscopes can use. And that's going to happen big time.
We're going to have a whole new understanding of cells,
not at the level of membranes running around, but of the individual
protein molecules running around each other.
Back up to what Tom was talking about, the death of electron
microscopy. That period when its imminent death seemed
unavoidable. There are a couple of other factors that I'd like
to bring up. One is that that was a period when light
microscopy took off, when it first started. And light
microscopy was providing two incredibly fundamental things that EM
did not provide. Number one, the dynamics of cells.
The actual movements of all of these things. Mind you,
light microscopy is 1/1000 the resolution, that far away.
But it was a whole process displayed for you in a living cell.
Second, the light microscopy was allowing you to begin to identify
which protein was doing it, because new technology of fluorescence,
fluorescent proteins and so on, were coming in to be examined by
light microscopy. So, it was not that electron microscopy died, it was
eclipsed by light microscopy and the huge potential of it.
You went through a light microscopy phase.
I fell in love with it myself, and lost myself in it.
But, the critical thing we've always been up against, and we're still
up against, no matter how fancy the electron microscopes get,
or how perfect our means of viewing is, we'll never get around
the fact that the electron microscope gives you a static
image. A still, a snapshot of something. One instant in time.
For each picture that you take, each preparation that you make,
is one instant in time. And you have to back extrapolate
to what was happening in life. You have to say, well I timed it
perfectly, I had 5 milliseconds, I had 10 milliseconds,
20 milliseconds, so I can say it did this, that, and that.
But that's an interpretation in and of itself.
Open to questions if you got competitors in your field.
It's always that extrapolation event that's an interpretation.
And that made people shy off and led to the demise,
of which we're now trying to recover. Always will be trying to
recover.
I mean, if you were a young student in the course and you came in
and you had a choice of spending a week doing meticulous
preparation to get something ready for the electron microscope
to get one picture of it, or you could take some genes and create some
beautiful colors that would run up and down inside your cell,
you could sit there all afternoon looking at them, which would you do?
Got me. I'd sit here and look at this. That's one picture.
So what is the future of electron microscopy then? Why
-- what recommends it to students?
Well, you asked two questions there. The future and
what recommends it to students. It might be that there's little to
recommend to the students. That's what we're really facing.
How many people are going into this field? It's heartbreaking to see
how few people are being trained, even in these labs that
are totally cutting edge. And the whole culture of electron microscopy
is in dire danger.
So still, you haven't answered why they should be trained,
what are they going to see that you can't see in a light
electron microscope?
Everything. I mean you're very limited. I mean the idea people
got all excited about high resolution microscopy and super
resolution. Super resolution, that's about a 100% improvement.
You go from seeing something that's 200 nm to something
that's 50nm. But we see stuff all the time that's 1nm!
2nm. I mean, the gap between resolution in electron
microscope and a light microscope is huge. And I don't
-- my goal in my current work is that I want to understand
how individual proteins fit together to make protein motors.
Now you can examine that with light microscopy, but
with some fluorescent dyes and watch what happens,
make some inferences, but you don't actually know until you
get the images and reconstruct the proteins.
This is -- I've just listened to Tom give his lectures
to this year's students at the Marine Biological Laboratory
and was impressed again by what an appearance he has.
He, as he's saying, and it's apparent in every word he says
to students, he believes, and I totally agree with him, that
it's at the EM resolution, a level of electron microscopy,
that we're going to understand cell function, organelle function,
and everything about the cell. Because it's at that level
that you can actually see molecules. And the whole name of the game
is seeing molecules interacting with each other.
That's -- to understand this machine called life, we have
to see the individual molecules interacting with each other.
Light microscopy gives you these wonderful clues that
they're dancing together, or going over here, but to see
how that machine works, you have to have the resolution of
this instrument. And it's a simple fact. It's amazing that we have
the instrument. It's great that we've had it for 50 years,
60 years almost, but it's got a niche in cell biology
and in medical science that is irreplaceable.
Never will be the same.