That depends on the projection used to obtain the map image.
This post is wrong from this point on.
If you draw a three points on the map such that
Ay == By && Ax != Bx
Ax == Cx && Ay != Cy
+=====================+
# #
# A-------------B #
# | #
# | #
# C #
# #
+=====================+
Is Alat == Blat && Along == Clong?
-
If so, every pixel represents the same long and lat difference.
Pixellat = (Blat - Alat) / (Bx - Ax)
Pixellong = (Clong - Along) / (Cy - Ay)
-
If not, the math gets a bit more complicated, but it's still pretty straightforward. You'll probably need a fourth point D such that
Dx == Bx && Dy == Cy
+=====================+
# #
# A-------------B #
# | | #
# | | #
# C-------------D #
# #
+=====================+
There surely exists modules to help you here.