diff --git a/CHANGELOG.md b/CHANGELOG.md
index ceb2b635..76ea8384 100644
--- a/CHANGELOG.md
+++ b/CHANGELOG.md
@@ -13,6 +13,7 @@
 
 ### Improvements
 * Added support for Numpy 2 and h5py 3.12, and pinned PyNWB to <3.0 temporarily. [#536](https://github.com/NeurodataWithoutBorders/nwbinspector/pull/536)
+* Added best practice around not using colons in object names. [#532](https://github.com/NeurodataWithoutBorders/nwbinspector/pull/532)
 
 ### Fixes
 * Fixed issue where the description check failed if the description was a list. [#535](https://github.com/NeurodataWithoutBorders/nwbinspector/pull/535)
diff --git a/docs/best_practices/general.rst b/docs/best_practices/general.rst
index f4ed6356..9c523047 100644
--- a/docs/best_practices/general.rst
+++ b/docs/best_practices/general.rst
@@ -34,15 +34,16 @@ It is recommended to use :wikipedia:`CamelCase <Camel_case>` when naming instanc
 
 
 .. _best_practice_name_slashes:
+.. _best_practice_name_colons:
 
-Do Not Use Slashes in Names
-~~~~~~~~~~~~~~~~~~~~~~~~~~~
+Do Not Use Slashes or Colons in Names
+~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
 
-Slash characters ``'/'`` and ``'\'``  should not be used in the ``name`` of an object, because they can be confusing to systems that parse HDF5 files (which NWB uses as the primary backend, see the :nwb-overview:`NWB FAQ <faq_details/why_hdf5.html#why-hdf5>` for more details). The forward slash is used in `h5py` to specify a `Groups <https://schema-language.readthedocs.io/en/latest/description.html#groups>`_ hierarchy.
+Slash characters ``'/'`` and ``'\'`` and colons ``':'`` should not be used in the ``name`` of an object, because they can be confusing to systems that parse HDF5 files (which NWB uses as the primary backend, see the :nwb-overview:`NWB FAQ <faq_details/why_hdf5.html#why-hdf5>` for more details) and/or Zarr files (which NWB allows as a backend). The forward slash is used in `h5py` to specify a `Groups <https://schema-language.readthedocs.io/en/latest/description.html#groups>`_ hierarchy.
 
 For mathematical expressions, instead of including the special character in the name, please use an English equivalent instead. *E.g.*, instead of "Df/f" use "DfOverF".
 
-Check function: :py:meth:`~nwbinspector.checks._general.check_name_slashes`
+Check functions: :py:meth:`~nwbinspector.checks._general.check_name_slashes` and :py:meth:`~nwbinspector.checks._general.check_name_colons`
 
 
 
diff --git a/src/nwbinspector/checks/__init__.py b/src/nwbinspector/checks/__init__.py
index a24e6323..5c3d39a0 100644
--- a/src/nwbinspector/checks/__init__.py
+++ b/src/nwbinspector/checks/__init__.py
@@ -12,6 +12,7 @@
 )
 from ._general import (
     check_description,
+    check_name_colons,
     check_name_slashes,
 )
 from ._icephys import (
@@ -91,6 +92,7 @@
     "check_spike_times_not_in_unobserved_interval",
     "check_description",
     "check_name_slashes",
+    "check_name_colons",
     "check_image_series_data_size",
     "check_image_series_external_file_relative",
     "check_image_series_external_file_valid",
diff --git a/src/nwbinspector/checks/_general.py b/src/nwbinspector/checks/_general.py
index 1eefd1a6..5aacbce6 100644
--- a/src/nwbinspector/checks/_general.py
+++ b/src/nwbinspector/checks/_general.py
@@ -16,6 +16,15 @@ def check_name_slashes(neurodata_object: object) -> Optional[InspectorMessage]:
     return None
 
 
+@register_check(importance=Importance.BEST_PRACTICE_SUGGESTION, neurodata_type=None)
+def check_name_colons(neurodata_object: object) -> Optional[InspectorMessage]:
+    """Check if an object name contains a colon."""
+    if hasattr(neurodata_object, "name") and ":" in neurodata_object.name:
+        return InspectorMessage(message="Object name contains colons.")
+
+    return None
+
+
 @register_check(importance=Importance.BEST_PRACTICE_SUGGESTION, neurodata_type=None)
 def check_description(neurodata_object: object) -> Optional[InspectorMessage]:
     """
diff --git a/tests/unit_tests/test_general.py b/tests/unit_tests/test_general.py
index d386ad61..6231bd29 100644
--- a/tests/unit_tests/test_general.py
+++ b/tests/unit_tests/test_general.py
@@ -1,7 +1,7 @@
 from hdmf.common import DynamicTable, DynamicTableRegion
 
 from nwbinspector import Importance, InspectorMessage
-from nwbinspector.checks import check_description, check_name_slashes
+from nwbinspector.checks import check_description, check_name_colons, check_name_slashes
 
 
 def test_check_name_slashes_pass():
@@ -10,17 +10,48 @@ def test_check_name_slashes_pass():
 
 
 def test_check_name_slashes_fail():
-    """HDMF/PyNWB forbid "/" in the object names. Might need an external file written in MATLAB to test that?"""
-    for x in ["\\"]:
-        table = DynamicTable(name=f"test{x}ing", description="")
-        assert check_name_slashes(neurodata_object=table) == InspectorMessage(
-            message="Object name contains slashes.",
-            importance=Importance.CRITICAL,
-            check_function_name="check_name_slashes",
-            object_type="DynamicTable",
-            object_name=f"test{x}ing",
-            location="/",
-        )
+    # the latest version of HDMF/PyNWB forbids "/" in the object names when creating a new object
+    # so we use in_construct_mode=True to simulate an object that was read from a file
+    table = DynamicTable.__new__(DynamicTable, in_construct_mode=True)
+    table.__init__(name="test/ing", description="")
+    assert check_name_slashes(neurodata_object=table) == InspectorMessage(
+        message="Object name contains slashes.",
+        importance=Importance.CRITICAL,
+        check_function_name="check_name_slashes",
+        object_type="DynamicTable",
+        object_name="test/ing",
+        location="/",
+    )
+
+    table = DynamicTable(name="test\\ing", description="")
+    assert check_name_slashes(neurodata_object=table) == InspectorMessage(
+        message="Object name contains slashes.",
+        importance=Importance.CRITICAL,
+        check_function_name="check_name_slashes",
+        object_type="DynamicTable",
+        object_name="test\\ing",
+        location="/",
+    )
+
+
+def test_check_name_colons_pass():
+    table = DynamicTable(name="test_name", description="")
+    assert check_name_colons(neurodata_object=table) is None
+
+
+def test_check_name_colons_fail():
+    # the latest version of HDMF/PyNWB forbids ":" in the object names when creating a new object
+    # so we use in_construct_mode=True to simulate an object that was read from a file
+    table = DynamicTable.__new__(DynamicTable, in_construct_mode=True)
+    table.__init__(name="test:ing", description="")
+    assert check_name_colons(neurodata_object=table) == InspectorMessage(
+        message="Object name contains colons.",
+        importance=Importance.BEST_PRACTICE_SUGGESTION,
+        check_function_name="check_name_colons",
+        object_type="DynamicTable",
+        object_name="test:ing",
+        location="/",
+    )
 
 
 def test_check_description_pass():